Fig. 6
Nuclear TRIM59 promotes macroH2A1 ubiquitination and degradation. a, b WB and qRT-PCR assays of effects of TRIM59 knockdown on macroH2A1 protein (a) and mRNA (b) levels in LN229/EGFR and U87/EGFR cells stimulated with EGF. c, e Effects of TRIM59 knockdown on macroH2A1 (mH2A1) stability in LN229/EGFR (c) and U87/EGFR (e) cells. After stimulation with EGF (100 ng/ml) for 6 h, cells were treated with cycloheximide (CHX, 20 mg/ml) for the indicated time points. d, f Quantification of mH2A1 protein levels in c and e, respectively. g Effects of TRIM59 WT, S308A, or S308D on mH2A1.1 ubiquitination. His-tagged ubiquitin (His-Ub) was co-transfected into LN229/EGFR/shTRIM59 and U87/EGFR/shTRIM59 cells with TRIM59 constructs or empty vector control. h Effects of Roscovitine on mH2A1.1 ubiquitination. His-Ub was co-transfected into LN229/EGFR/shTRIM59 and U87/EGFR/shTRIM59 cells with TRIM59 constructs or empty vector control. i HA-TRIM59 proteins purified from HEK293 cells were incubated with ATP, E1, and E2 along with Flag-mH2A1.1 proteins isolated from HEK293 cells for in vitro ubiquitination assay. Data are representative of three independent experiments with similar results. Data were expressed as means ± SD. **P < 0.01 by two-tailed Student’s t test. Source data are provided as a Source Data file
