FIGURE 2.
FM19G11 regulates Oct4 and Sox2 pluripotency markers in rodent and human stem cells. epSPC (A and B) and hESC (C): A, upper panel: real time SYBR Green PCR analysis of rat Sox2 and Oct4 relative expression levels. Lower panel, semi-quantitative PCR of Oct4, Sox2, and their direct target genes, Tgfα and Nanog, respectively. Cells were incubated with 500 nm FM19G11 (+) or its vehicle (−) for 48 h under hypoxia (1% O2). 20% O2 was taken as the basal condition. GAPDH served as a loading control. B, left panels: representative Western blot (upper panels) and densitometry analysis of three independent experiments (lower panels). FM19G11 dose-dependent effect on Oct4 and Sox2 protein expression after 48 h of hypoxia exposure. Values are shown as a percentage of the control (20% O2). Right panels: representative immunostaining of the Sox2 protein in undifferentiated neurospheres treated for 48 h in 1% O2 with vehicle or 500 nm FM19G11. C, left panel: TaqMan real time-PCR analysis of human Sox2 and Oct4 relative expression levels in hESC treated with 500 nm FM19G11 or its vehicle; right panel: qualitative immunostaining analysis of human Sox2 protein expression in undifferentiated hESC colonies treated for 48 h in 1% O2 with DMSO (vehicle) or 500 nm FM19G11. 20% O2 condition served as the basal control. Results were obtained from three independent experiments. Error bars represent S.D. *, p < 0.05 versus 20% O2; §, p < 0.05 versus vehicle at 1% O2, determined by Student's t test.