Cell culture and transfection reagents

All cell culture and transfection reagents were from Invitrogen; and standard reagents were from Sigma-Aldrich or Fisher Scientific. Drugs were from: PLX4032/GSK1120212/PD0325901/Erlotinib (Selleck); and FCCP, Antimycin A, 4-OHT, mDIVI-1 (Sigma-Aldrich). Antibodies: DRP1^Total, DRP1^S616Ⓟ, DRP1^S637Ⓟ, ERK^Total, ERK^Ⓟ (Cell Signaling); RAS (EMD/Millipore Anti pan RAS AB-3); Mfn2 (Abcam); OPA1 (BD Biosciences); Actin, GAPDH, HSP60, Mfn1, ND1, ND2, SMAC (Santa Cruz). MitoTracker Green and Hoechst 33342 are from Invitrogen and Anaspec, respectively.

All cell culture and transfection reagents were from Invitrogen; and standard reagents were from Sigma or Fisher Scientific. Drugs were from: ABT-737 (Abbott Pharmaceuticals), ABT-263/PLX-4032/GSK-110212 (Selleck), zVAD-fmk (Calbiochem), Hoechst 33342 (Anaspec), and staurosporine/cisplatin/dacarbazine/vinblastine (Sigma). Antibodies (clone): anti-actin (C4), anti-A1 (FL-175), anti-BCL-2 (100), anti-CD31 (BD Pharmingen #550274) anti-MCL-1 (Rockland), anti-BAD (C7), anti-BIM (22 (link)–40), anti-PUMA (CT; Sigma), anti-SMAC (H177), anti-BID (C20), anti-cytochrome c (7H8.2C12), anti-BAK (G23), anti-BAX (6A7 for IP; N20 for western blot), anti-BCL-xL (H5 for IP; S18 for western blot), anti-GAPDH (9B3), anti-HSP60 (B-9), anti-p44/p42 MAPK ERK1/2 (137F5), and anti-phospho p44/42 MAPK ERK1/2 (197G2). Caspase-8 cleaved mouse BID (C8-BID) was from R&D Systems. Full-length BAX was made as described (24 (link)). Human BCL-xLΔC, MCL-1ΔC, and PUMAβ were made as described (17 (link)). The human BIM BH3 domain peptide (> 98% purity, Abgent) was resuspended in anhydrous DMSO in a N₂ environment, stored at −80°C, and thawed only once. All lipids for the LUVs studies were purchased from Avanti Polar Lipids. Statistical significance was evaluated by two tailed Student t test for p < 0.05.

All cell culture and transfection reagents were from Invitrogen; and standard reagents were from Sigma or Fisher Scientific unless indicated. Drugs were from: ABT-737 (Abbott Pharmaceuticals), Hoechst 33342 (Anaspec); and β-ME, Cisplatin, DTT, JC-1, mDIVI-1, Paclitaxel, Tg, TMRE, and Tun (Sigma). Antibodies (clone): anti-actin (C4), anti-BCL-2 (100), anti-MCL-1 (Rockland), anti-BIM (22-40), anti-PUMA (CT; Sigma), anti-cytochrome c (7H8.2C12), anti-BAK (NT), anti-BAX (clone 6A7 for IP; clone N20 for western blot; clone Δ21 for trypsin studies), anti-BCL-xL (S18), anti-GAPDH (9B3), anti-Mfn1 (ABCAM, 57602), anti-Mfn2 (ABCAM, 56889), anti-HSP60 (B-9). BAX, BAX^ΔC, BAX^S184A, BCL-xL^ΔC, N/C-BID, BIM-S were made as described (Chipuk et al., 2008 (link); Suzuki et al., 2000 (link); von Ahsen et al., 2000 (link)). The human BIM and PUMA BH3 domain peptides (Abgent) were resuspended in anhydrous DMSO. Knockout and Wt matched MEFs: Bak^−/−, Bax^−/−, Bak^−/−Bax^−/−, Bid^−/−, Bid^−/−Bim^−/−, Puma^−/−, Mfn1^−/− and Mfn2^−/− were obtained from Drs. Stanley Korsmeyer (Wei et al., 2001 (link)), Doug Green (Chipuk et al., 2008 (link)), Gerald Zambetti (Jeffers et al., 2003 (link)), and ATCC (Chen et al., 2003 (link)).