Fig. 2

ERK1/2 regulates the H3K9me2 and DNA methylation reprogramming in paternal genomes. A H3K9me2 staining of control, 50 μM GDC-0994-treated, and 100 μM GDC-0994-treated zygotes at PN4-5 stage (10 hpf). B Values are represented as the mean of H3K9me2 signal intensity in paternal and maternal pronuclei after background subtraction (left axis) or a ratio between signals of parental pronuclei (pat./mat., right axis). Number of zygotes analysed for each group: control n = 10; 50 μM GDC-0094-treated n = 15; 100 μM GDC-0994-treated n = 15. C 5mC (green) and 5caC (red) staining of control, 50 μM GDC-0994-treated, and 100 μM GDC-0994-treated zygotes at PN4-5 stage (10 hpf). D, E Quantification of 5mC and 5caC is represented as signal intensity in paternal and maternal pronuclei (left axis) or as a ratio between parental signals (pat./mat., right axis). Number of zygotes analysed for each group: control n = 23; 50 μM GDC-0994-treated n = 34; 100 μM GDC-0994-treated n = 28. Statistical analysis was carried out using Student’s t-test (two-sided). P values are indicated. Error bars indicate SD. ♀, maternal pronucleus; ♂, paternal pronucleus. Scale bar, 20 µm