Figure 4

Ubiquitinated and SUMOylated SETD2 after proteasome inhibition by bortezomib in HMC-1 cells (a) and in SM patients (b). Co-immunoprecipitation (IP) with an anti-SETD2 antibody and immunoblotting (western blotting) with anti-ubiquitin and anti-SUMO antibodies performed before and after bortezomib treatment (10 nM for 24 h). H3K36Me3 levels were also assessed by WB. H3 and beta-actin were used as controls. The two ISM patients had near-normal SETD2 protein levels (90.5% and 75.8% of SETD2 expression with respect to a pool of healthy donors); the MCL patient had no SETD2 protein expression (see Supplementary Table 3 for SETD2 protein level estimates as assessed by densitometric analysis of western blots). One of three independent experiments is shown in all panels.