Figure 6.

BRG1 regulates GIC sensitivity to TMZ. (A): GBM6 and X16 GICs were treated with 300 or 500 μM of TMZ, and cell proliferation was determined by CellTiter-Glo assays. (B): Total RNA was prepared from GBM6, X10 and X16 GICs, and MGMT expression was determined by qPCR. (C): Protein lysates from GBM6, X10 and X16 GICs were immunoblotted for MGMT and actin. (D): Control and BRG1-KD X16 GICs were treated with or without 500 μM of TMZ, and cell proliferation was determined by CellTiter-Glo assays. (E): Protein lysates from control (scrambled shRNA) and BRG1-KD GICs were immunoblotted for MGMT and actin. (F): The effect of TMZ on the tumorigenicity of control and BRG-KD GICs was assessed by injection of 10⁶ tumor cells into the flanks of NSG mice and live animal imaging was performed at weekly intervals. After confirming tumor initiation by live animal imaging, mice were treated with TMZ by IP injection (10 mg/kg on alternate days), and the reduction in tumor volume was calculated and 1 and 2 weeks post-treatment. (G) Tumor weights and (H) photographs of tumors at necropsy. *p ≤ .05 was considered to be statistically significant.