Fig. 2

Rhox6 relies on Nanos3 to enhance the formation of PGCLCs. A Western blot analysis of the expression of Flag and Rhox6 in i-Rhox6 mESCs, in which Flag-tagged Rhox6 was driven by the Tet-On system, in the presence or absence of 2 μg/ml Dox. B qRT-PCR analysis of the expression of Rhox6 and PGC marker genes in i-Rhox6 mESCs treated with Dox for 6 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. Wo, without; wi, with. C qRT-PCR analysis of the expression of Rhox6 and PGC marker genes in 46C mESCs infected with scramble or Rhox6 shRNA lentiviruses. The data are presented as the mean ± SD (N = 3 biological replicates). *P < 0.05, **P < 0.01 versus scramble, as determined by one-way ANOVA with Sidak’s multiple comparisons test. D Western blot analysis of the expression levels of Flag and Nanos3 in PB and PB-Rhox6 mESCs infected with scramble or Nanos3 shRNA lentiviruses. E qRT-PCR was used to detect the expression level of Nanos3 in and PB-Rhox6 mESCs infected with scramble or Nanos3 shRNA lentiviruses. The data are presented as the mean ± SD (N = 3 biological replicates). **P < 0.01 versus PB/scramble, as determined by one-way ANOVA with Sidak’s multiple comparisons test. F qRT-PCR analysis of the expression of PGC marker genes in PB and PB-Rhox6 PGCLCs infected with scramble or Nanos3 shRNA lentiviruses. The data are presented as the mean ± SD (N = 3 biological replicates). *P < 0.05, **P < 0.01 versus PB/scramble, as determined by one-way ANOVA with Sidak’s multiple comparisons test. G Western blot assay of the expression levels of HA, Rhox6 and Nanos3 in PB and PB-Nanos3 mESCs infected with scramble or Rhox6 shRNA lentiviruses. H qRT-PCR analysis of the expression levels of Rhox6 in PB and PB-Nanos3 mESCs infected with scramble or Rhox6 shRNA lentiviruses. The data are presented as the mean ± SD (N = 3 biological replicates). **P < 0.01 versus PB/scramble, as determined by one-way ANOVA with Sidak’s multiple comparisons test. I qRT-PCR analysis of the expression of PGC genes in PB and PB-Nanos3 PGCLCs infected with scramble or Rhox6 shRNA lentiviruses. The data are presented as the mean ± SD (N = 3 biological replicates). *P < 0.05, **P < 0.01 versus PB/scramble, ^#P < 0.05, ^##P < 0.01 versus PB/Rhox6 sh#1, ^&&P < 0.01 versus PB/Rhox6 sh#2, as determined by one-way ANOVA with Sidak’s multiple comparisons test