Figure 6: Characterization of hiPSCs generated using AxPou2.
From: Reprogramming to pluripotency is an ancient trait of vertebrate Oct4 and Pou2 proteins
(a) Immunofluorescence analysis of NANOG, OCT4 and SSEA4 on iPS1 aSK and iPS2 aSKM cell lines. OSK iPSC cell line was used as a positive control. Nuclei were stained with Hoechst (blue). Scale bars, 500 μm. (b) Expression of endogenous pluripotency markers in hFib1, iPS1 aSK, iPS1 aSKM, iPS2 aSK and iPS2 aSKM cell lines as well as in Shef3 hESCs was measured by qRT–PCR and plotted relative to Shef3 hESC levels. Error bars reflect s.e. based on normalization to GAPDH and ACTB (housekeeping genes). (c) Expression level of the viral transgenes in hiPSCs was measured by qRT–PCR using specific primers. Fibroblasts collected 4 days after infection are used for comparison as a positive control. Error bars represent the s.e. arising from using GAPDH and ACTB for normalization. (d) In vitro differentiation of the AxPou2 hiPSCs into cells of all three germ layers as shown by immunocytochemistry: endoderm (α-fetoprotein, AFP), mesoderm (α-smooth muscle actin, SMA), and ectoderm (β-Tubulin IIIb, TUJ1). Nuclei were stained with Hoechst (blue). Scale bars, 250 μm.
