Fig. 2

β-TrCP controls ARID1A protein levels. a NCI-N87 cells were transfected with con-shRNA or shRNA specific against β-TrCP for 36 h, cell extracts were subjected to western blot with the indicated antibodies. b AGS cells were transfected with con-shRNA or shRNA specific against β-TrCP for 36 h, cell extracts were subjected to western blot with the indicated antibodies. c NCI-N87 cells were transfected with con-shRNA or shRNA specific against β-TrCP for 24 h, and treated with 2 μg/ml VP16 for 6 h, 25 μg/ml (cycloheximide) CHX was added for the indicated time course. Cell extracts were subjected to western blot with the indicated antibodies. d NCI-N87 cells were co-transfected with both Flag-ARID1A and HA-β-TrCP plasmids for 24 h, and then treated with or without 2 μg/ml VP16 for additional 6 h. Cell extracts were IP with anti-HA antibody, and the immunocomplexes were subjected to western blot with the indicated antibodies. e NCI-N87 cells were transfected with Flag-ARID1A and his-ubiquitin with or without HA-β-TrCP plasmids for 24 h, and then treated with or without 2 μg/ml VP16 for additional 6 h, 20 μM MG132 was added 4 h before cell harvest. Cell extracts were purified with Ni⁺ beads, and the immunocomplexes were subjected to western blot with the indicated antibodies