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. 2020 Mar 4;8:103. doi: 10.3389/fcell.2020.00103

FIGURE 5.

FIGURE 5

Ectopic DNMT3L interacted with the PRC2 member SUZ12. (A) DNM3L or RFP expression was induced by 48 h of treatment with doxycycline (DOX) in lentivirus-infected DOX-inducible MEFs expressing the manufactured gene (Supplementary Figure S6). Western blot (WB) analysis of extracts of the same numbers of DNMT3L-expressing- (iD3L + DOX) and red fluorescent protein (RFP)-expressing (iRFP + DOX) MEFs using anti-DNMT3L antibody. Anti-GAPDH antibody was used as a loading control. (B,C) Immunoprecipitation (IP) and WB results revealed the interaction between DNMT3L and PRC2 members. Whole-cell extracts from DNMT3L- and RFP-expressing MEFs at 48 h postinduction were subjected to IP with anti–FLAG and anti–SUZ12 antibodies. WB using antibodies against DNMT3L, SUZ12, and EZH2 was performed to detect these proteins in the indicated IP-pulldown lysate extracts. SUZ12 and EZH2 showed equal quantities after a 5% input of both DNMT3L- and RFP-expressing MEFs. The presence of DNMT3L and SUZ12 was found in each protein interaction complex.