Supplemental Fig. 1 from Familial and Somatic BAP1 Mutations Inactivate ASXL1/2-Mediated Allosteric Regulation of BAP1 Deubiquitinase by Targeting Multiple Independent Domains

The deletion R666-H669 in the ULD domain of BAP1 corresponding to that observed in a uveal melanoma completely abolished ASXL2 binding in vivo. HEK 293 cells were co-transiently transfected with plasmids expressing WT or mutant BAP1 and ASXL2 (261-649aa) proteins. At 48 h post-transfection, the cells were collected and whole cell lysates were used for co-IP with ANTI-Flag M2 Affinity Gel. Western blot analyses were performed with BAP1 (rAb) and Flag (rAb).