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Fig. 6 | Cell & Bioscience

Fig. 6

From: Regulation of paternal 5mC oxidation and H3K9me2 asymmetry by ERK1/2 in mouse zygotes

Fig. 6

ERK1/2 inhibition promotes Tet3-driven oxidation of de novo 5mC in paternal pronuclei. A Impediment of de novo 5mC by Decitabine affects paternal 5hmC accumulation in ERK1/2-inhibited zygotes (10 hpf) as assessed by 5mC and 5hmC staining. B Quantification of 5mC (left axis) and 5hmC (right axis) is represented as the signal intensity in paternal pronuclei (n = 18 for control, n = 17 for iERK1/2, n = 17 for iERK1/2 + iDNMT; n = 21 for iDNMT). C Tet3 staining under PT conditions of the control, 50 μM GDC-0094-treated, and 100 μM GDC-0994-treated zygotes at PN3 stage (5.5 hpf). D Quantification of Tet3 is represented as signal intensity in paternal pronuclei after background subtraction. Number of zygotes analysed for each group: control n = 13, 50 μM GDC-0994-treated n = 14, 100 μM GDC-0994-treated n = 15. E Inhibition of Tet enzymes by DMOG impedes the paternal 5hmC accumulation ERK inhibition produced (50 μM GDC). Representative images of 5hmC and 5mC staining at PN4-5 (10 hpf). F Quantification of 5mC (left axis) and 5hmC (left axis) is represented as signal intensity in paternal pronuclei (n = 23 for control, n = 22 for iERK1/2, n = 26 for iERK1/2 + iTet3; n = 22 for iTet3). 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

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