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Fig. 2 | Clinical Epigenetics

Fig. 2

From: Differential effect of histone H3.3 depletion on retroviral repression in embryonic stem cells

Fig. 2

Effect of H3.3 depletion on proviral expression. A Sanger sequencing of the first coding exon of H3f3b gene following Crispr/Cas9 transfection and sorting of clones. B Following infection with lentiviral vector caring antisense H3f3a shRNA, full depletion of H3.3 was verified by RT-qPCR, normalized to UBC control gene. Data are the mean ± s.e.m. (n = 3) C Viral copy number calibration of PBSpro/PBSgln MLV pseudovirus. Levels of viral insertion into the cell genome were determined and normalized to NIH3T3 cells carrying a single integration site. D The percentage of GFP-positive after infection by PBSpro or E PBSgln virus in the H3.3 depleted ESCs, normalized to the WT PBSpro GFP expression. The red asterisk shows a significant difference from WT ESC (One sample Wilcoxon test), and the difference between the cells before and after the restoration of H3.3 expression is denoted by a black asterisk. Data are the mean ± s.e.m. (n = 5) F GFP expression levels of H3.3 depleted ESC, infected with PBSpro or G PBSgln MLV virus, days after infection (dai). All P values are calculated using a two-tailed unpaired Mann–Whitney U test; Data are the mean ± s.e.m.*P < 0.05, **P < 0.01

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