Figure 7. Pou3f1 alleviates the inhibitory effects of BMP4 and Wnt3a on neural fate commitment.
(A) Inducible Pou3f1-overexpressing ESCs were cultured as EBs in serum-free medium for 4 days with or without BMP4/Dox treatment from day 2 to day 4. Gene expression levels were detected by Q-PCR. (B) Immunocytochemical assays using day 4 EBs described in A. The EBs were stained with Sox (red) and with Oct4 (green). Scale bars, 100 μm. (C) Statistical analysis of results from the immunocytochemical assay of Sox+/Oct4−, Nestin+, and Tuj1+ cells in EBs and of Tuj1+ replated cells. (D) Pou3f1-overexpressing ESCs were cultured as EBs in serum-free medium for 4 days with or without Wnt3a/Dox addition from day 2 to day 4. Gene expression levels were detected by Q-PCR. (E) Pou3f1 partially rescues the inhibitory effects of xBMP4 on cSox2. In situ hybridization of cSox2 (blue) in chick embryos that were co-electroporated with xBMP4 plus IRES-GFP (control vector, a and a′) or Pou3f1-IRES-GFP (b and b′), respectively. GFP expression (brown) was detected in a′ and b′ by immunohistochemistry. The values represent the mean ± SD for A, C, and D. (*p<0.05; **p<0.01).