Effect of FASN on TNF-α expression. A, D: Effect of FASN on basal TNF-α expression. Endogenous TNF-α mRNA was determined using real-time RT-PCR assay of total RNA from stable MCF7 cells with FASN overexpression (FASN1 and FASN2) and vector-transfected control (Vec) cells (A) and stable MCF7/AdVp3000 cells with FASN knockdown (Si) or cells transfected with scrambled control shRNA (Scr) (D). B, C, E, F: Effect of FASN on drug induced TNF-α expression. TNF-α mRNA and protein were determined using real-time RT-PCR (B, E, F) and ELISA (C) assays in stable MCF7 cells with FASN overexpression (FASN1 and FASN2) and vector-transfected control (Vec) cells following treatment with 1 µM doxorubicin (B, C) and in stable MCF7/AdVp3000 cells with FASN knockdown (Si) or transfected with scrambled control shRNA (Scr) following treatment with 1 µM doxorubicin (E) or 10 µM mitoxantrone (F) in the presence of 10 µM FTC. DOX-induced TNF-α protein increase represents the net increase of TNF-α protein following doxorubicin treatment over that of the untreated control cells. **P < 0.01.