Fig. 2

Gas5 is required for mESC self-renewal. a, b Differentiating morphological change was observed in Gas5 knockdown (KD) mouse embryonic stem cells (mESCs). c The endogenous expression of Gas5 expression in Activin A- or retinoic acid-induced differentiation of mESCs. d The pluripotency marker ALP staining in Gas5 KD mESCs. e Gas5 expression in six important pluripotent transcriptional factors Nanog, Sox2, Oct4, Tcl1, Esrrb, and Tet1 KD mESCs. f mRNA expression of six pluripotent transcriptional factors (TFs) Nanog, Sox2, Oct4, Tcl1, Esrrb, and Tet1 in Gas5 KD mESCs. g Protein expression of Sox2 and Oct4 in Gas5 KD mESCs. h Gas5 expression during a time-elapsed knockdown assay over a period of 8 days following either Sox2, Oct4, Nanog, Esrrb, and Tcl1 KD in mESCs. i Chromatin immunoprecipitation (ChIP) analysis of the binding of Oct4, Sox2, and cMyc to Gas5 promoter by using Oct4, Sox2, and cMyc antibodies. *P < 0.05, **P < 0.01, ***P < 0.001, by t test, n = 3. Error bars represent SEM of the indicated experiment replicates. GFP green fluorescent protein, lncRNA long noncoding RNA, mLIF mouse leukemia inhibitory factor, shRNA short hairpin RNA