Figure 4

Reduced Mule/Mcl-1 complex detected in breast cancer cells compared with mammary epithelial cells. (A) Mammary epithelial cells (MCF-10A and HMLE, left panel) and breast cancer cells (MDA-MB-468, MDA-MB-231 and MCF-7) were treated with CHX (200 μg ml^–1) for various time points (0–24 h) and 100 μg of protein lysates were immunoblotted using anti-Mule antibody. (B) (Top panel) Mammary epithelial cells (MCF-10A and HMLE, left panel) and breast cancer cells (MCF-7 and MDA-MB-468, right panel) were treated with CHX (200 μg ml^–1) for various time points (0–36 h) and immunoprecipitated with anti-Mule antibody or with IgG (16 h) and immunoblotted for Mcl-1. (Bottom panel) MCF-10A cells were treated with CHX (200 μg ml^–1) for 0–36 h and immunoprecipitated with anti-Mule antibody or with IgG (16 h) and immunoblotted for Bcl-2. (C) Mammary epithelial cells (MCF-10A and HMLE, left panel) and breast cancer cells (MCF-7 and MDA-MB-468) were treated with CHX (200 μg ml^–1) for various time points (0–36 h) and immunoprecipitated with anti-Mcl-1 antibody or with IgG (16 h) and immunoblotted for Mule. (D) (Top panel) Small-inhibitory RNA (siRNA)-mediated inhibition of Mule in mammary epithelial cells (HMLE and MCF-10A). Western blot analysis showing Mule (top) and GAPDH (bottom). (Bottom panel) Measurement of apoptotic index in random (Ran) and Mule siRNA (Mule siRNA) treated mammary epithelial cells by TUNEL assay after CHX treatment. ^**P<0.01.