To do this, you'll need to go to Fortnite's login page and create a new account using either email or your second PlayStation Network account. If the second player doesn't have a Fortnite account or doesn't want to complete the linking process, you'll need to create a separate Epic account to link to your console. In this tutorial video I will show you how to setup a split screen configuration for any game on PC, Xbox, Playstation or any console.
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Hair regrowth and thickening treatments, shampoos, styling products and supplements are formulated to address hair loss and thinning, helping to encourage healthy growth with over-the-counter medicinal, cosmeceutical and natural ingredients. If hair and scalp concerns are getting in the way of you feeling and looking your best, you can discover products to address your issues in our selection of hair care products. You'll find all of these styling aids and more here at Walgreens. Finish things off with hairspray to secure every strand. What it is: An argan oil-infused, anti-frizz hairspray with moveable but firm hold.Gels make it easier to sculpt and shape and you can count on serums to smooth away frizz in a flash.Add texture with pomade, wax and other texturizers or increase body and bounce with mousse and volumizing sprays.Fortunately, styling products can improve manageability, making it easier to create all types of hairstyles. Solutions for Your Styling ProblemsĮven if you wash and condition on a regular basis, your hair may not be easy to style and hold in place. ![]() Salon shampoos and conditioners allow you to purify and hydrate with the same products trusted by the pros. You can choose to purchase shampoo and conditioner individually or as a part of hair care sets that give you the convenience of one-and-done shopping. We carry shampoo and conditioner for all hair types and textures, and with products designed specifically for children and ethic hair types, we offer something for everyone. Choose from Same Day Delivery, Drive Up or Order Pickup plus free shipping on orders 35+. ![]() Shop Target for moroccan oil you will love at great low prices. Use throughout the day to shield from UV damage.While hair types differ greatly, all hair requires regular cleansing with shampoo and moisturizing with conditioner to remain in the best of health. Garnier Whole Blends Illuminating Leave In Conditioner Moroccan Argan & Camellia Oils - 5.1 fl oz. Finish by misting all cover for optimal shine and to control frizz. Spray Organix Renewing Moroccan Argan Oil Healing Oil evenly onto damp hair then blow dry to infuse it into the hair shafts. This rare formula has a centuries-old history of providing natural vitamin E and antioxidants while renewing your hair's cell structure, sealing in shine, and creating lush softness.Īfter shampooing, towel dry hair.This exotic precious formula of Moroccan Argan Oil Weightless spray instantly heals split ends, creating soft frizz-free perfection while protecting your hair from harmful styling heat and UV damage, giving a smooth, glossy, shine.It's the perfect indulgence for soft, seductive, silky perfection!.This exotic precious formula of Moroccan argan oil spray instantly infuses hair with a sheer veil of shine, giving color and highlights a multifaceted glow, while it mends split ends, and tames fly-a-way frizzies. Same great formula in new packaging! Image & details will be revised, shortly) Read honest and unbiased product reviews from our users. (Product received may temporarily differ from image shown due to product update. Find helpful customer reviews and review ratings for Moroccanoil Luminous Hairspray, Extra Strong, Travel Size, 75ml at. The price is high, but so is the performance. Combine that with superb bokeh, near perfect control of chromatic aberration, and minimal distortion, and you've got a killer all-around lens. ![]() It's a little soft even in the center at f/1.2, but it quickly comes into its own and by f/4 is possibly the sharpest lens you can get in Fuji's system. The Fujinon XF 56mm f/1.2 R isn't a perfect lens, but it's certainly on the right track. A perfect lens would render the finest details accurately, wouldn’t distort straight lines or produce ugly fringing around high-contrast subjects, and would create smooth out-of-focus areas. When evaluating any lens, we focus on four key areas: sharpness, distortion, chromatic aberration, and bokeh. The only Fuji owners who won’t have this lens at the very top of their wishlists will be people who already own it. There are sharper lenses wide open, and there are definitely cheaper ones, but this is a superb all-around performer that Fuji can be very proud of. In the end, the Fujinon 56mm f/1.2 R is one of the best examples of a classic portrait lens that you’ll find for any mirrorless system. The major drawback is slow autofocus, but if you're shooting someone sitting for a portrait, that's basically a non-issue. Of course, if you don't need razor-thin depth of field you can also consider the super-sharp XF 60mm f/2.4 R Macro, which also doubles as an excellent portrait lens and costs just $550 at major retailers. ![]() On top of the $1,000 you’ll pay for the “vanilla” version, that's a pretty tall order. On the downside, it doesn’t work with phase-detect autofocus and costs $500 more. You can find samples from this lens online, but the major difference is that it produces even softer, better-rounded bokeh. ![]() Just about the only competition this lens has in Fuji's system is its own APD variant. The ultra-wide f/1.2 aperture creates some amazing bokeh. It's the best way to make your subject pop, ensuring all eyes are This kind of focal length helps to isolate your subject, effectively compressing the background while blurring it away with shallow depth of field. Designed for the APS-C sensors used in Fuji's X-series cameras, it has a full-frame equivalent focal length of 84mm to go with its f/1.2 max aperture-a classic combo. The Fujinon 56mm f/1.2 R is a wide-aperture, medium telephoto prime lens that's designed primarily for portrait photography. It proved its mettle in our labs, but with a $1,000 asking price, is it worth the cost? Who's It For? This ultra-bright prime lens features a classic portrait focal length and one of the widest apertures you can get in an autofocus lens. ![]() While typically quite expensive, these lenses serve as "halo" products, showcasing what each manufacturer's system is capable of.įujifilm's halo lens is undoubtedly the Fujinon XF 56mm f/1.2 R (MSRP $999.99). Lenses like the Carl Zeiss 24mm f/1.4 for Sony E-mount and Panasonic 42.5mm f/1.2 Nocticron for Micro Four Thirds stand out for their exceptional sharpness and beautiful bokeh. While many are designed to be as small and affordable as possible, a few excel in the pursuit of pure photographic performance. The latter group have made a smart decision by opting to craft high-quality prime lenses in lieu of more beginner-friendly zooms. ![]() A fee is charged for use of these stations. RV Sanitary Stations are available at the Sideling Hill, Allentown, Lawn and Valley Forge service plazas. In the app, to the right of the map, tap the 'Your Location' option. It’s drawn from domestically drilled natural gas wells or in conjunction with crude oil production.ĬNG is available at the New Stanton service plaza. To start, open the Google Maps app on your phone. Consisting mostly of methane, CNG is odorless, colorless and tasteless. Compressed Natural Gas (CNG)Ĭompressed Natural Gas (CNG) is a readily available alternative to gasoline that’s made by compressing natural gas to less than 1% of its volume at standard atmospheric pressure. Caltrans provides Safety Roadside Rest Areas as a part of the State Highway System pursuant to Streets and Highways Code, Sections 218-226.5. E-85 fuelĮ-85 fuel is available at the Oakmont Plum service plaza. ![]() In addition, Tesla Electric Superchargers are available at North and South Somerset, Bowmansville, Hickory Run, and Peter J. See the Blink Network website for current charges. Customers can pay using a credit card through the Blink App, or by contacting Blink at 1-88. The charging stations are located in the car parking lots. New Stanton, King of Prussia - Blink Level II and Level III Units. With decades of experience in innovative transportation software technology, TRT's founders, Dayton Males and Thomas Bouve, ensure top-notch. Looking for directions to the nearest rest area Developed by Trip Routing Technologies, LLC (TRT), our patented and ad-free app prioritizes enhanced travel tools for drivers and highway service providers. Or even better, join a local guide program to share your favorite stores.Electric vehicle charging stations are installed and ready for customers to use at five turnpike service plazas: Oakmont Plum, Bowmansville, Peter J. Navigate hassle-free with Rest Stops Ahead. Torn between two businesses? You can start by looking at comments and photos shared by locals to make a more informed decision. Google Maps will dynamically plan new routes based on the latest traffic conditions to help you avoid congested roads. ![]() And because Google Maps reroutes based on real-time traffic, drivers can complete trips faster and get more rides in. Drivers can rely on the Google Maps experience they’re used to, including intuitive turn-by-turn navigation, route overviews, lane level guidance, and voice support. Instead of having to switch back and forth between apps, drivers efficiently control their queue and see passenger info all in one place, making it easier to manage rides and get to where they’re going. Give your drivers seamless experiences by integrating turn-by-turn navigation powered by Google Maps right into your app. Highway 20 Rest Area is a 2 acre area that is a popular stopping spot for. Help your drivers get from point A to Z faster and safer Skip Navigation Links Home > Counties > Calhoun > Parks > Highway 20 Rest Area. ![]() Using the Map To make the map easier to use as you travel, you can turn layers on and off within the map legend. You can see everything from the highest peak to the lowest canyon. The map is interactive so you can zoom in closer for more detail, select a marker for rest area information, or select the View larger map icon at top-right to open the map in Google Maps. If a map is no longer just a map, what is it? It’s a trip to the moon, a ticket to Mars, a bird's eye view of the Earth. Revisit the world around you in ways you've never seen before Express your appreciation for the place you like with a comment, express your dissatisfaction with the place you don't like with a score, and add your own photos and record every place you go on your trip. Papa T’s homemade Italian sausage links broiled with peppers, onions and mushrooms. Chesters Fried Chicken Food 0.23 mi away. Chicken Wings, Burgers, Sandwiches 0.16 mi away. deliveries.Even in the 1990s when the Bills were Superballs almost-champions I paid a waiger to a Dallas friend with La Nova wings. La Nova Pizzeria « Back To Springdale, PA. A mix of Romaine & Iceberg Lettuce, Tomatoes, Grated Cheese, Croutons & Anchovies (on request) Small 8.49. Chicken ParmigianaĬhicken cutlets breaded and sautéed covered with mozzarella cheese and baked until golden brown and topped with our homemade red sauce, served over spaghetti View the online menu of La Nova Pizzeria and other restaurants in Springdale, Pennsylvania. Your choice of 5 cold subs, served on a platter.Īll Entree Orders Served with a Sheet of Garlic Toast, Napkins, Plates. Get La Nova Italian Seasoned Cooked Chicken Wings delivered to you in as fast as 1 hour via Instacart or choose curbside or in-store pickup. Enter your address above to see fees, and delivery + pickup estimates. Romaine Lettuce Topped with Chicken, Tomatoes, Onions, Sweet Peppers, Celery, Black Olives, Croutons and Feta Cheese ![]() Romaine Lettuce topped with Ham, Salami, Capicola, Tomatoes, Onion, Celery, Chi-Chi Beans, Black Olives, Green Olives, Cheddar Cheese, Pepperoni, Banana Peppers ![]() Overnight Shipping is included in the cost for the wings (mild, medium, hot or char-BQ) which can be delivered in the lower 48 and as far away as Alaska. Half (Serves 8-10) $52.40 | Whole (Serves 18-20) $75.33Īdd grilled chicken to any salad – Half $13.09 | Whole $21.82 The wings the Daily Meal called America’s Best & named one of America’s best regional foods would be a huge hit at your Bills game party. La Nova Pizzeria - Chicken wings No Blue Cheese tasted like kerosene. Romaine Lettuce topped with Tomatoes, Onions, Romano Cheese, Croutons and anchovies (on request) The wings tasted like they had been read more. Romaine Lettuce topped with Tomatoes, Black Olives, Green Olives, Onion, Celery, Chi-Chi Beans, Sweet Peppers and Croutons ![]() Prices include overnight shipping costs for. Topped with La Nova chicken fingers with four different cheeses. This meaty, moist wing has sold millions La Nova wings are Frozen. $14.04 ($18.37 w/Sauce) Salads Chef Salad The flavor known around the world as 'Buffalo Style' These wings are covered in our famous hot sauce recipe with just the right blend of spices to them the true Original Buffalo Wings Flavor'. Williamsville Location (view updated menu)ĥ151 Main Street Williamsville, NY 14222 Appetizers Garlic Knots ![]() In a separate essay, Post music critic Chris Richards reminds us not to forget the past 20 years as we celebrate hip-hop’s 50th. You might, of course, encounter explicit lyrics. Listen to the 50 songs as you scroll, by toggling the sound on and off, or enjoy them as a Spotify playlist here. By having artists share what they love about hip-hop - its beauty, its insight, its unapologetic nature - we hope they’ll remind you of what you love too, or introduce you to songs you’d overlooked. ![]() In short, we passed the mic to hip-hop and let it freestyle. We talked to hip-hop figures from various eras and regions, including a few non-artists, looking for anything but standard answers. With that in mind, we asked people who made the music we love to tell us about songs they love - the ones that still make them feel the way they first did, or those that inspired, impacted or influenced them. But no top 50 can encompass all that this form of cultural expression has become, or include everyone who has played a part in it. in the Bronx, widely regarded as the birth of hip-hop - we could have made a ranked list you’d argue about. 11 approaches - 50 years from the day DJ Kool Herc threw a 1973 back-to-school party for his sister Cindy at 1520 Sedgwick Ave. Ip-hop is an art form that rewards style and originality yet constantly pays homage - through samples, shout-outs and collaborations - to the artists on whose shoulders it stands. Complete your look with black leather derby shoes to instantly kick up the fashion factor of this look. Here are our Top 10 Favorite Ways to Style Light Blue Jeans: Form-fitting sweatshirt with a t-shirt peeking out underneath Collared undershirt with sweater. This pairing of a tan trenchcoat and light blue jeans can only be described as ridiculously sharp and casually classy. Blue jeans can be paired with tank tops, formal shirts tucked in or out, T-shirts, button-downs for work, peplum tops or long tops for a modest look. Give casualness to your outfit by finishing with a pair of white leather low top sneakers. Teaming a grey overcoat with light blue jeans is a good option for a casually neat ensemble. When it comes to shoes, this outfit is finished off perfectly with tan suede slip-on sneakers.įavorite Unfavorite White Crew-neck T-shirt Going for a yellow button-up as opposed to a short-sleeved alternative is a quick and easy way to dress up this combination. The dark blue jeans perfectly contrast with the jacket and make you look like a million bucks. Items: Mustard Yellow Button-Up Shirt, Dark Blue Jeans, Tan Brown Dress Shoes, Light Grey Blazer. The jacket is made of soft, supple leather that will keep you warm all night long. Light Blue Jeans Outfit Inspiration: What To Wear With Pale Denim. This combination of a light blue denim shirt jacket and light blue jeans is on the casual side but is also stylish and incredibly stylish. The brown leather jacket with dark blue jeans for men is the perfect outfit for a night out. Tone down the casualness of this look with black leather loafers.įavorite Unfavorite Orange Crew-neck T-shirt Lightweight Reversible Seamless Stretchable. To add more class to your look, round off with brown leather chelsea boots.įavorite Unfavorite Black Crew-neck T-shirtĪ red and white varsity jacket and light blue jeans are indispensable menswear must-haves if you're planning a casual wardrobe that holds to the highest fashion standards. XUGWLKJ Womens Blue Denim Dress Blouses Long Sleeve Big Swing Casual Dresses. ![]() Light Blue Vertical Striped Long Sleeve Shirtĭress in a white short sleeve shirt and light blue jeans to pull together a casual and cool ensemble. ![]() What To Wear With Light Wash Jeans 1.Sign up to see recommended looks Get started Favorite Unfavorite White Short Sleeve Shirt There are 15 different looks in this article so there is bound to be one that suits your look, or you can play around with one of these outfits to really make it your own! Unique Bargains Womens Plus Size Outfits Babydoll Denim V Neck Short Sleeve Midi. In this article, we will give you some tips on what to wear with light wash jeans or light blue jeans for the most stylish outcome. Light wash denim jeans can be dressed up or down depending on what you wear them with, which makes them perfect for lots of different occasions ranging from a casual weekend look to evenings or dinner out! They are perfect for summer (well, all seasons really!), and there are so many different ways to style them, no matter your body type or size. Looking for a fun and stylish way to dress up your new jeans? Here’s a style guide on what to wear with light wash jeans for a super chic look.ĭo you have a pair of light wash jeans in your closet? If not, you’re missing out on a key fashion staple that is incredibly versatile! We may earn a commission if you decide to make a purchase through our links, at no cost to you. See more ideas about light blue jeans outfit, light blue jeans, outfits. Pair them with light blue ripped and cuffed skinny. Explore Yofashionqueen32s board 'Light blue jeans outfit' on Pinterest. ![]() We only feature products that we hope you will love. One of the top jean trends, light wash jeans have been seen on those in the fashion know for quite a while now. To form this warm and attractive outfit, you can simple wear a white t shirt with a grey sweater cardigan. ![]() (Truthfully, though, Darren was a much better Man-At-Arms Darren grew up to be a doctor despite his debilitating fear of blood so playing the part of the brilliant engineer ended up being way more in his wheelhouse than a muscular hero who occasionally scraped his knee.)Īs a self-professed tomboy, during recess I was always playing He-Man with my guy friends. Jesse was always He-Man, except on the rare occasion that Darren Young tried to take over the role. When it came to our hierarchy, Jesse Correia was He-Man. ![]() On our playground, though, we were all devoted He-Man fans and that was that. Though no new episodes of He-Man and the Masters of the Universe were being developed, She-Ra: Princess of Power was slowly gaining ground.     Masters of the Universe: Revelation - He-Man and playground politicsĭespite the complexities of kindergarten playground politics in the mid-1980s, it was a universally accepted truth at Loma Portal Elementary that if you were going to “play He-Man” at recess, it was a co-ed experience.īy that point, the cartoon had ended its run and we were all watching reruns. So why was I so conflicted about it? To answer this question, I had to go back to where it all began. He-Man is tied to some of my happiest memories as a child, and I’d been so excited to see it come back to television. He died soon thereafter, leaving behind a brilliant legacy tainted by shoddy business deals that ultimately robbed him of the ability to cash in on the franchise he helped build. I was fortunate enough to meet Filmation’s Lou Scheimer at San Diego Comic-Con, twice, and I cried at the panel when he announced it would be his last public appearance. The vitriol aimed at the new series was on my mind all weekend, long after I’d watched all five episodes.Īs a 1980s child, I grew up with He-Man. Despite not hating it, though, I realized I had complicated feelings about the new series that I just couldn’t shake. It felt like an old-school episode of He-Man. I loved the animation and the music and the cast and the writing. But I didn’t expect the anger from a large swath of fans who saw it as an attempt to make it the Teela show. I figured there would be some backlash from the people you can’t please no matter what you do, because such is the nature of fandoms these days. Smith, a lifelong nerd who understands the importance of protecting beloved characters and franchises, was the perfect person to bring the series to life. Not only was my favorite childhood hero returning, but Kevin Smith was behind it. If some of your content was shared by accident.The day that Netflix announced Masters of the Universe: Revelation was a happy day for me. We do our best to keep tabs on infringements. Entertainment News International is not responsible for reporting errors, inaccuracies, omissions, and or other liablities related to news shared here. All content sourced by fans, online websites, and or other fan community sources. ![]() Data has been shared for news reporting purposes only. ©Entertainment News International - All images, trademarks, logos, video, brands and images used on this website are registered trademarks of their respective companies and owners. Get the scoop on all the popular comics, games, movies, toys, and more every day! Contact and Support Entertainment News International (ENI) is the #1 popular culture network for adult fans all around the world. ![]() For information on how to obtain certain costumes, see Skin Catalogue. Gallery consisting of artworks used primarily in-game. The first 200 K31s were made in May 1931 for troop trials (serials 500,001 – 500,200), which is where the '31' in the rifle's name originates. Model 1889 Rifle - Antique - NO FFL REQUIRED - Surplus Good/Very Good Condition. The Karabiner Model 1931 was a new design created by the Eidgenössische Waffenfabrik in Bern, Switzerland under Colonel Adolf Furrer (1873–1958). Neither was mechanical engineer Eduard Rubin (1846–1920), the designer of the 7.5×55mm Swiss ammunition previous Swiss service rifles and the K31 were chambered for. This is a beautiful example of a Swiss Model K31 rifle manufactured in 1947. This rifle was manufactured in 1947, and comes with the original Swiss leather sling. It was the standard issue rifle of the Swiss armed forces from 1933 until 1958. The K31 began to be gradually replaced by the Stgw 57 beginning in 1958.Īlthough the K31 is a straight-pull design broadly based on previous Swiss "Schmidt–Rubin" service rifles and carbines, the K31 was not designed by Colonel Rudolf Schmidt (1832–1898), as he was not alive in 1931 to do so. The Karabiner Model 1931 (K31) is a magazine-fed, straight-pull bolt-action rifle. The Karabiner Model 1931 replaced the Model 1911 rifle and carbine in Swiss military service, and would serve from the early 1930's through the Second World War and into the Cold War. ![]() A charger is used to load the magazine from the top of the receiver. implement work meaning Switzerland Karabiner Model 1931 (k31) - For Sale :: K31: The Swiss Rifle That my like new 1958 K31 - Swiss Rifles dot com. It has a 6-round removable magazine, and is chambered for the 7. It was the standard issue rifle of the Swiss armed forces from 1933 until 1958 though examples remained in service into the 1970s. It has a 6-round removable magazine, and is chambered for the 7.5×55mm Swiss Gewehrpatrone 1911 cartridge (aka GP 11). It was the standard issue rifle of the Swiss armed forces from 1933 until 1958, though examples remained in service into the 1970s. Each rifle included a 6-round detachable box magazine with matching stamped serial number. 31) is a magazine-fed, straight-pull bolt-action rifle. The Karabiner Model 1931 (K31) is a magazine-fed, straight-pull bolt action rifle. The K31 is chambered for the 7.5×55mm Swiss Gewehrpatrone 1911 or GP 11 cartridge, a full-power rifle cartridge with ballistic qualities similar to the 7.62×51mm NATO/.308 Winchester cartridge. The Karabiner Model 1931 (K31) is a magazinefed, straightpull bolt action rifle. The original rifles were massive and had strange, elongated receivers fitted with 12. The original designs for the gun were adopted into service in 1889, just as Switzerland entered the era of smokeless powder. It was the standard issue rifle of the Swiss armed forces from 1933 until 1958 though examples remained in service into the 1970s. The K31 Schmidt-Rubin Short Rifle (Karabiner Model 1931) is a very unique design that dates back to the blackpowder cartridge days. The Karabiner Model 1931 (K31) is a magazine-fed, straight-pull bolt action rifle. ![]() ![]() hPGCLCs are aggregated with mouse embryonic ovarian somatic cells to form xenogeneic reconstituted ovaries, which are cultured under an air–liquid interface condition for ~4 months for hPGCLCs to differentiate into oogonia and immediate precursory states for oocytes. iMeLCs, or, alternatively, hPSCs cultured with divergent signaling inhibitors, are induced into hPGC-like cells (hPGCLCs) in floating aggregates by cytokines including bone morphogenic protein 4. hiPSCs are induced into incipient mesoderm-like cells (iMeLCs) using activin A and a WNT pathway agonist. Here, we describe a protocol to differentiate human induced pluripotent stem cells (hiPSCs) into oogonia in vitro. hPGCs undergo genome-wide epigenetic reprogramming and differentiate into oogonia or gonocytes, precursors for oocytes or spermatogonia, respectively. (I) Amplitude, maximal upstroke velocity (Vmax upstroke), and maximal decay velocity (Vmax decay) of calcium transients.The human germ-cell lineage originates as human primordial germ cells (hPGCs). (H) Representative tracings of rhythmic spontaneous and caffeine-induced calcium transients. Bottom: action potential duration at 90 and 50% repolarization (APD90 and APD50). (G) Top: maximal diastolic potential (MDP). Data correspond to clone UC C0406-iPS-C4P17 (also in G through I) similar results were observed with UC C0406-iPS-C1P17. (F) Representative spontaneous action potential tracing recorded by whole-cell patch clamp technique. ASGPR stands for asialoglycoprotein receptor. Glycogen accumulation was detected with periodic acid-Schiff staining. (D and E) Phase contrast and immunofluorescence photographs of hepatocytes and cardiomyocytes produced from representative UiPSC clones scale bars, 50 μm (phase contrast of cardiomyocytes), 200 μm (phase contrast of hepatocytes) and 50 μm (all immunofluorescence photographs). Bottom: confocal immunofluorescence of neurons and astrocytes (glial fibrillary acidic protein ) produced from the same UiPSC clone scale bars, 50 μm. Middle: confocal immunofluorescence microscopy for the indicated markers of neural rosettes produced from the same UiPSC clone scale bars, 50 μm. (C) Top, from left to right: phase contrast photographs of EBs growing in suspension, neural rosettes, and neurons produced from a representative UiPSC clone scale bars, 50 μm. AFP stands for α-fetoprotein scale bars, 50 μm. (B) Confocal immunofluorescence microscopy for markers of the 3 germ layers in differentiating embryoid bodies (EBs) of a representative UiPSC clone. (A) Teratomas comprising derivatives of the 3 germ layers. Multidifferentiation potential of urinary induced pluripotent stem cells (UiPSCs). (J) Bisulfite sequencing analysis for the Oct4 and Nanog proximal promoters in 2 representative UiPSC clones from the same donor. (I) Normal karyotype of representative male and female UiPSC clones. (H) Semiquantitative PCR showing integration of the exogenous transgenes in the genome of the indicated UiPSC clones, urine donor cells, water, and H9 ESCs were included as controls. (G) Scatter plot of DNA microarrays data for 2 representative UiPSC clones and H9 ESCs. (F) qPCR showing silencing of the exogenous transgenes in the indicated UiPSC clones values are referred to transduced cells extracted at day 6. Values are referred to donor urine cells H9 ESCs were used as control. hTERT indicates human telomerase reverse transcriptase. (E) Quantitative real-time PCR (qPCR) for endogenous ESC transcription factors in the indicated UiPSCs. (D) Confocal immunofluorescence microscopy for the indicated human embryonic stem cell (ESC) markers of a representative UiPSC clone scale bars, 100 μm. Bottom: representative phase contrast photographs of passage (P) 1 UiPSCs grown on feeders (alkaline phosphatase staining is also included) or Matrigel. (C) Top: representative phase contrast photographs of emerging urinary induced pluripotent stem cells (UiPSCs) colonies at different time points. Cells infected with the exogenous factors are also included note the early morphologic changes (clustering) indicative of reprogramming. (B) Phase contrast and immunofluorescence photographs of urine cells at day 3 after infection with control green fluorescent protein (GFP) retrovirus. SKOM refers to the four exogenous factors Sox2, Klf4, Oct4, and c-Myc. (A) Schematic representation of iPSC generation from urine cells (UC). Generation of induced pluripotent stem cells (iPSCs) from urine cells. |
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