Fig. 4

ERK1/2 contributes to the asymmetry of parental H3K9me2 by preventing paternal G9a localization. A G9a staining of control, 50 μM and 100 μM GDC-0994-treated zygotes at PN3 stage (5.5 hpf). B Quantification is represented as the mean of G9a signal intensity on parental pronuclei (left axis) or a ratio of G9a signal intensity between parental pronuclei (pat./mat., right axis). Number of zygotes analysed for each group: control n = 14; 50 μM GDC-0994-treated n = 26; 100 μM GDC-0994-treated n = 16. C Inhibition of G9a enzymes by BIX-01294 affects the deposition of paternal H3K9me2 at PN4-5 stage (10 hpf). D Quantification of the H3K9me2 signal intensity in parental pronuclei (left axis) or a ratio between the pronuclei signal (pat./mat., right axis). For H3K9me2 staining, control n = 29, iERK1/2 (100 μM GDC-0994) n = 19, iERK1/2 + iG9a n = 21; iG9a n = 14. E Under the same treatment conditions, G9a staining of control, iERK1/2 (100 μM GDC-0994), iERK1/2 + iG9a, and iG9a zygotes at PN4-5 stage (10 hpf). F Quantification of G9a staining in both paternal and maternal pronuclei (left axis) or a ratio between the pronuclear signals (pat./mat., right axis). For G9a staining, control n = 27, iERK1/2 n = 34, iERK1/2 + iG9a n = 40; iG9a 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. PB, polar body. Scale bar, 20 µm