Fig. 3

Ssbp3 depletion weakens the trophoblast gene expression induced by BMP4 and bFGF treatment in ESCs. a The morphology of E14T cells after treatment with bone BMP4 and bFGF at the indicated time points. b Expression levels of Ssbp3 gradually increased in E14T cells treated with BMP4 and bFGF. The expression levels of Ssbp3 were determined by qRT-PCR analysis. The average mRNA level in E14T cells cultured without treatment was set at 1.0. Data are shown as mean ± SD (n = 3). *p < 0.05, **p < 0.01. c The silencing efficiency of shRNAs targeting Ssbp3 in E14T cells was detected by qRT-PCR analysis at day 4. d–i The expression levels of trophoblast-specific markers in cells expressing NT control shRNA or specific Ssbp3-targeting shRNAs were determined by qRT-PCR analysis. E14T cells were induced to trophoblast differentiation by treatment with BMP4 and bFGF for 96 h. The average mRNA level in shNT-expressing E14T cells without treatment was set at 1.0. Data are shown as mean ± SD (n = 3). *p < 0.05, **p < 0.01. bFGF basic fibroblast growth factor, BMP4 bone morphogenetic protein 4, NT non-targeting, shRNA short-hairpin RNA