Fig. 1

FBW7 negatively regulates the stability of Brg1. a Immunoblot analysis (IB) of whole cell lysates (WCLs) derived from wild-type (WT) and FBW7^−/− DLD1 cells. b, c WT and FBW7^−/− DLD1 cells were treated with 20 μg/ml cycloheximide (CHX). At the indicated time points, WCLs were prepared and IB analysis was carried out with indicated antibodies (b). The relative Brg1 intensity was normalized to Vinculin and then normalized to the t = 0 controls (c). d IB analysis of WCLs derived from WT and FBW7^−/− DLD1 cells treated with MG132 (10 μM) or DMSO for 10 h. e IB analysis of WCLs derived from MKN45 cells infected with the indicated shRNA lentiviruses. f IB analysis of WCLs derived from MKN45 cells infected with the indicated shRNA lentiviruses and treated with 20 μg/ml CHX for indicated time periods. The relative intensity of Brg1 was normalized to Vinculin and then normalized to the t = 0 control (Bottom). g IB analysis of WCLs and immunoprecipitated (IPs) derived from 293T cells transfected with Flag-Brg1 together with the indicated constructs of Myc-tagged Cullin family members. h IB analysis of WCLs derived from MKN45 cells infected with the indicated lentiviral shRNA-Cullin1 constructs. i IB analysis of WCLs and IPs derived from 293T cells transfected with Flag-Brg1 together with the indicated FBW7 constructs. j Co-IP experiments in MKN45 cells were performed using anti-Brg1 antibody (sc-17796, Santa Cruz). Mouse IgG was used as a control. k IB analysis of WCLs derived from FBW7^−/− DLD1 cells infected with the indicated FBW7-expressing lentiviral vectors