Irdye anti rabbit or anti mouse secondary antibodies

For general immunoblotting, cells were lysed in 1% Nonidet P-40 (NP-40) lysis buffer (50 mM Tris [pH 7.4], 150 mM NaCl, 0.5 mM EDTA, 1% NP-40, 0.5% sodium deoxycholate, 1 mM Na₃VO₄, 1 mM NaF) + complete protease inhibitors (Roche) for 30 min on ice. Lysates were cleared by centrifugation, boiled in 2x sample buffer (Laemmli buffer + 50 μM 2-βME) and separated on 4–20% SDS-PAGE gels (Bio-Rad). Proteins were transferred to nitrocellulose membranes for 10 min using the TransBlot Turbo system (Bio-Rad) and subsequently blocked with Odyssey blocking buffer (LI-COR). Blots were probed with the following Abs: anti-FASN (Cell Signaling Technology #3180), anti-MCL-1 (Cell Signaling Technology #94296), anti-BCL-2 (Cell Signaling Technology #4223), anti-FASL (BD Pharminogen #556372), anti-BIM (Enzo Life Sciences), anti-NUR77 (Biolegend, clone 1E10A15) and anti-β-actin (Sigma-Aldrich). Blots were incubated with IRDye anti-Rabbit or anti-Mouse secondary antibodies (LI-COR) and imaged using the Odyssey CLx instrument. Band intensity quantification was conducted with StudioLite Software (LI-COR).