MSLN overexpression makes PC cells resistant to TNF-α induced apoptosis through reduced Caspase 3 cleavage and up regulated anti-apoptotic proteins. (A). Reduced Caspase 3 cleavage in MIA-MSLN cells by 20 ng/ml of TNF-α treatment for 72 h. (B). Silencing endogenous MSLN in AsPC-3 cells renders it sensitive to TNF-α induced apoptosis through increased Caspase 3 cleavage. Panel (i) shows the reduction of MSLN expression level in AsPC-shMSLN stable cell line. Panel (ii) shows the increased caspase 3 cleavage upon TNF-α treatment. (C). Pre-treatment with pan-caspase inhibitor zVAD-fmk renders MIA-V cells resistant to TNF-α-induced apoptosis. Cells were treated with 20 ng/ml of TNF-α for 72 h and then stained with TUNEL assay. (D). Pro and anti-apoptotic molecules expression in MIA-V/MIA-MSLN cells with or without treatment of 20 ng/ml TNF-α assayed by Western blot. (E). Effect of different TNF-α doses on selected pro/anti-apoptotic molecules expression in MIA-V/MIA-MSLN cells. (F). Effect of zVAD-fmk pre-treatment on various pro and anti-apoptotic molecules expression in MIA-V/MIA-MSLN cells after treatment with 50 ng/ml of TNF-α for 72 h.