Changes in histone modification marks and nucleosome dynamics upon Sox8 promoter Wnt signaling activation. (A and B) Levels of repression of mark H3K27me3 (A) and activation of mark H3K4me3 (B) at the Tcf4 binding site (bp −800) and the mrhl RNA binding site (bp −141) on the Sox8 promoter in Gc1-Spg cells (control and Wnt3A conditioned medium treated) and mouse testes (P7 and P21). (C) Micrococcal nuclease digestion of chromatin for different time intervals (50, 55, and 60 min) from control and Wnt3A conditioned medium-treated Gc1-Spg cells to obtain mononucleosomes. (D) Change in nucleosome dynamics on the 1-kb upstream Sox8 promoter upon Wnt signaling activation. Numbers 1 to 19 represent every 50-bp interval scored for in the 1-kb upstream Sox8 gene promoter, with 19 being adjacent to the TSS. The black line represents the Tcf4 binding site, and the red line represents chromatin oligonucleotide affinity purification (ChOP) read results. Data are plotted as means ± SD, n = 4. ***, P ≤ 0.001; **, P ≤ 0.01; *, P ≤ 0.05 (two-tailed Student t test). N. S, not significant.