Fig. 3

Nanos3 is a direct target of Rhox6. A CUT&Tag analysis of the enrichment of Rhox6 in the promoters of Blimp1, Tfap2c, Prdm14, Nanos3, Stella and Mup1. B The Nanos3 promoter (from − 2000 to + 1) was used as a template, and 10 pairs of qRT-PCR primers were designed. C ChIP assays were performed using an anti-Flag antibody. IgG was used as a negative control. The fold enrichment in the indicated regions of the Nanos3 promoter was measured by qRT-PCR. The data are presented as the mean ± SD (N = 3 biological replicates). **P < 0.01 versus IgG, as determined by Student’s t test. D Consensus binding motif of Rhox6 predicted by the AnimalTFDB database. E The binding position and sequence of Rhox6 in the Nanos3 promoter and the corresponding deletion mutation sequence. TSS, transcription start site. F Luciferase activity analysis of the WT or mutant (Mut) Nanos3 promoter reporter plasmid-expressing cell lines transfected with PB or PB-Rhox6. PB/Nanos3^WT was used as the control for normalization. The data are presented as the mean ± SD (N = 3 biological replicates). *P < 0.05, **P < 0.01 versus PB/Nanos3^WT, ^##P < 0.01 versus PB-Rhox6/Nanos3^WT, as determined by one-way ANOVA with Sidak’s multiple comparisons test. ns, not significant