Overexpression of GCNF inhibits Oct4 expression.
A, Western blot results of the level of GCNF and Oct4 proteins in undifferentiated G-hES cells treated with Dox (0.0, 0.125, 0.25, 0.5, 1.0, 2.0 μg/ml) for 4 days. β-Actin was used as a loading control. B, Western blot results of GCNF, Flag tag GCNF, Oct4, and β-actin expression in undifferentiated G-hES cell treated with Dox and without Dox for 4 days. C, Oct4 mRNAs were detected in undifferentiated G-hES cells, Dox-treated G-hES cells, RA-treated G-hES cells, and Dox-plus-RA treated G-hES cells. D, Sox2 mRNAs were detected in undifferentiated G-hES cells, Dox-treated G-hES cells, RA-treated G-hES cells, and Dox-plus-RA treated G-hES cells. E, Western blot results of GCNF and Oct4 expression in undifferentiated G-hES treated with Dox for 4 days, and in differentiated G-hES cells treated with RA or plus Dox for 2, 4, and 6 days. F, qRT-PCR results of GCNF expression in undifferentiated G-hES treated with Dox for 4 days and in differentiated G-hES cells treated with RA or plus Dox for 2, 4, and 6 days. G, qRT-PCR results of Oct4 mRNA expression in undifferentiated G-hES treated with Dox for 4 days and in differentiated G-hES cells treated with RA or plus Dox for 2, 4, and 6 days. * indicates no statistically significance with p ≥ 0.05; ** indicates statistically significance with p < 0.05.