Figure 2. BRG1 deficiency activates transcription of a subset of NRF2 target genes.

BRG1 and/or BRM were knocked down in H358 cell line using a lentiviral system or transfection to deliver shRNA. The A427 and H522 lung cancer cell lines possessing inactivating mutations of BRG1 and epigenetic silencing of BRM were used as controls. (A) The indicated human NSCLC cell line was transfected with either a non-targeting siRNA (siCONTROL) or a siRNA targeting BRG1 (siBRG1). 72h after transfection, the indicated proteins were detected and quantified by western blot analysis from whole cell lysates. Relative protein abundance was quantified by densitometry from linear-range film exposures. (B) mRNA levels were determined by qPCR for each gene normalized to B-ACTIN. *P-value< 0.05, (Student T test) and error bars represent ± SEM. qPCR results are representative of two independent experiments and assayed twice. (C) Whole cell lysates were separated by 4–12% SDS-PAGE and probed with the indicated antibodies. (D) Clonal H358 cells stably expressing BRG1 and/or BRM shRNAs were treated with tBHQ for 16h before protein extraction and western blot analysis. Protein abundance was quantified by densitometry from linear-range film exposures. B-actin-normalized values are presented. (E-J) Transcript abundance for the indicated mRNAs was determined by qPCR. B-ACTIN was used for normalization. Data represent two independent experiments, assayed in technical duplicate. * P-value < 0.05 (Student T test) and error bars represent ± SEM.