Fig. 1

Rhox6 promotes the differentiation of PGCLCs derived from mESCs. A Heatmap analysis of the expression of Rhox6, Rhox9 and PGC marker genes in mouse epiblasts and E9.5 PGCs. B qRT-PCR analysis of the expression of PGC marker genes and Rhox family members in mESCs, EpiLCs and PGCLCs. The data are presented as the mean ± SD (N = 3 biological replicates). *P < 0.05, **P < 0.01 versus mESCs, as determined by one-way ANOVA with Sidak’s multiple comparisons test. C Western blot analysis of the expression level of Flag in 46C mESCs transfected with PB, PB-Rhox6 or PB-Rhox9. β-Tubulin was used as the loading control. D qRT-PCR detected the expression of the PGC marker genes Prdm14, Nanos3 and Stella in PB and PB-Rhox9 PGCLCs. The data are presented as the mean ± SD (N = 3 biological replicates). **P < 0.01 versus PB, as determined by Student’s t test. E Immunofluorescence staining of Tfap2c expression in PB and PB-Rhox6 PGCLCs. Scale bar, 100 μM. F qRT-PCR analysis of the expression of PGC marker genes in PB and PB-Rhox6 PGCLCs. The data are presented as the mean ± SD (N = 3 biological replicates). **P < 0.01 versus PB, as determined by Student’s t test. G Flow cytometry analysis of the expression of Blimp1-mCherry in PB and PB-Rhox6 PGCLCs. H qRT-PCR analysis of the expression level of Rhox6 in 46C mESCs infected with scramble or Rhox6 shRNA lentiviruses. 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. I qRT-PCR analysis of the expression of PGC marker genes in scramble or Rhox6 shRNA PGCLCs. 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. J Immunofluorescence staining of Tfap2c expression in scramble and Rhox6 shRNA PGCLCs. Scale bar, 100 μM