Fig. 2

Ssbp3 depletion attenuates the activation of trophoblast gene expression induced by downregulation of Oct4 in mouse ESCs. a The morphology of ZHBTc4 cells after treatment with Tc. Differentiation was triggered by Tc-mediated downregulation of Oct4. b Expression levels of Ssbp3 during differentiation of the ZHBTc4 cell line were determined by qRT-PCR analysis. The average mRNA level in ZHBTc4 cells cultured without Tc was set at 1.0. Data are shown as mean ± SD (n = 3). *p < 0.05, **p < 0.01. c Protein levels of Ssbp3 and Oct4 during ZHBTc4 cell differentiation were determined by Western blotting. d Western blot analysis of the silencing efficiency of shRNAs targeting the Ssbp3 coding sequence in ZHBTc4 cells. e The expression levels of trophoblast-specific markers in cells expressing NT control shRNA or specific Ssbp3-targeting shRNA were determined by qRT-PCR analysis. Expression of Oct4 was downregulated by treatment with Tc for 96 h in ZHBTc4 ESCs. The average mRNA level in shNT-expressing ZHBTc4 cells cultured in the absence of Tc was set at 1.0. Data are shown as mean ± SD (n = 3). *p < 0.05, **p < 0.01. NT non-targeting, shRNA short-hairpin RNA, Tc tetracycline