Fig. 4.

H3K9me2 is reduced at the proviral LTR in G9a^−/− mESCs. (A) Depletion of H3K9me2 in G9a^−/− cells, but not in Suv39h1/2^−/− cells, was confirmed by Western blot analysis. ChIP was conducted on MSCV-GFP–infected TT2 and G9a^−/− mESCs using H3K9me2- and H3K9me3-specific antibodies or IgG as a control. (B) qPCR was conducted using primers specific for the endogenous Mage-a2 gene. The mean level of enrichment of technical replicates [expressed as a percentage (± SD) of input] is shown. (C) Amplification with primers specific for the MSCV 5′ LTR and GFP regions revealed a decrease in H3K9me2 at the 5′ LTR in the G9a^−/− line. (D) ChIP was conducted with H3K4me2- and H3K9ac-specific antibodies and analyzed by qPCR, as above.