Fig. 5

Screening of DEGs regulated by Rhox6 knockdown. A Volcano plot showing DEGs mediated by Rhox6 downregulation. B KEGG analysis of DEGs regulated by Rhox6 downregulation. C Heatmap analysis of the expression of pluripotency-associated genes in DEGs. D qRT-PCR analysis of the expression of candidate genes regulated by Rhox6 knockdown in C. The data are presented as the mean ± SD (N = 3 biological replicates). **P < 0.01 versus scramble, as determined by one-way ANOVA with Sidak’s multiple comparisons test. E qRT-PCR analysis of the expression of Rhox6 and the indicated genes in i-Rhox6 mESCs treated with or without Dox for 12 h. The data are presented as the mean ± SD (N = 3 biological replicates). **P < 0.01 versus wo Dox, as determined by Student’s t test. F qRT-PCR analysis of the expression of Id3, Wnt6, Klf4, Tbx3 and Lefty1 in WT and Rhox6-knockout mESCs. The data are presented as the mean ± SD (N = 3 biological replicates). **P < 0.01 versus Rhox6^WT, as determined by Student’s t test. G Western blot analysis of the protein levels of Flag and Tbx3 in i-Rhox6 mESCs treated with or without Dox for 12 h