Fig. 5

H3K27me3 was not recovered in oocytes after treatment of fetal ovaries with tazemetostat. Data summary from flow cytometric analyses of E11.5 XX gonads cultured for 24 h with DMSO (green), 100 nM or 1 μM tazemetostat (Blue), followed by drug removal and additional culture for 48 h in the presence of DMSO (green), 100 nM or 1 μM tazemetostat. Drugs were removed by washes with culture media after the initial 24 h of culture. Four treatment groups were used to collect control and experimental data. Treatment 1: DMSO/DMSO—24 h DMSO followed by 48 h DMSO (negative control). Treatment 2: TAZ—24 h tazemetostat (H3K27me3 depletion after 24 h drug treatment). Treatment 3: TAZ/DMSO—24 h tazemetostat followed by 48 h DMSO (H3K27me3 recovery after 48 h drug withdrawal). Treatment 4: TAZ/TAZ—24 h tazemetostat followed by 48 h tazemetostat (H3K27me3 depletion after 72 h drug treatment). a Summary of experimental pipeline for assessing H3K27me3 recovery after drug depletion in germ cells undergoing epigenetic reprogramming. b Average H3K27me3 staining intensity in germ cells for treatments 1–4 DMSO/DMSO, TAZ, TAZ/DMSO and TAZ/TAZ using 1 μM (left) and 100 nM (right) tazemetostat from cultured E11.5 XX female gonads (1 μM, n = 5 biological replicates, 100 nM n = 4 biological replicates). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; one-way ANOVA plus post hoc Tukey’s multiple comparisons test. Error bars ± SEM