Anti cyto c

For immunoblotting, samples resolved by SDS-PAGE were transferred onto 0.2 μm nitrocellulose membranes (Bio-Rad, cat# 1620112). Membranes were blocked with 5% non-fat milk, and then incubated with the appropriate primary antibodies: anti-HA (Sigma-Aldrich, cat# H3663), 1:5000; anti-Flag (Sigma-Aldrich, cat# F1804), 1:5000; anti-ICDH (Xu et al., 2010 (link)), 1:10,000; anti-tubulin (DSHB, E7) 1:10,000; anti-PDHA1 (ProteinTech, cat# 18068-1-AP), 1:5000; anti-ATPB (ProteinTech, cat# 17247-1-AP), 1:2000; anti-TOM20 (ProteinTech, cat# 11802-1-AP), 1:5000; anti-VDAC1 (ProteinTech, cat# 55259-1-AP), 1:2000; anti-Cyto c (Santa Cruz Biotechnology, cat# sc-13560), 1:1000; anti-Calnexin (ProteinTech, cat# 10427-2-AP), 1:2000; anti-ANT1/2 (ProteinTech, cat# 17796-1-AP), 1:1000; anti-GM130 (BD Biosciences, cat# 610822), 1:2500; and anti-ADPR (Sigma-Aldrich, cat# MABE1016), 1:1000. Membranes were then incubated with an appropriate IRDye infrared secondary antibody and scanned by using an Odyssey infrared imaging system (Li-Cor’s Biosciences).

Purified His₆‐Larg1 was used to raise mouse‐specific antibodies using a standard protocol (Pocono Rabbit Farm & Laboratory). The antibodies were affinity‐purified as previously described
⁶⁵ (link). For immunoblotting, samples resolved by SDS‐PAGE were transferred onto 0.2 μm nitrocellulose membranes (cat# 1620112; Bio‐Rad), which were blocked with 5% nonfat milk at room temperature for 1 h before being incubated with the appropriate primary antibodies: anti‐Flag (cat# F1804; Sigma‐Aldrich), 1: 5000; anti‐HA (cat# H3663; Sigma‐Aldrich), 1:5000; anti‐tubulin (E7; DSHB) 1:10,000; anti‐ADPR (cat# MABE1016; Sigma‐Aldrich), 1:1000; anti‐ICDH
⁶⁶ (link), 1:10,000; anti‐PDHA1 (cat# 18068‐1‐AP; Proteintech), 1:5000; anti‐ATPB (cat# 17247‐1‐AP; Proteintech), 1:2000; anti‐TOM20 (cat# 11802‐1‐AP; Proteintech), 1:5000; anti‐VDAC1 (cat# 55259‐1‐AP; Proteintech), 1:2000; anti‐Cyto‐c (cat# sc‐13560; Santa Cruz), 1:1000; anti‐Larg1, 1:2500. Membranes were then incubated with appropriate IRDye infrared secondary antibodies and scanned by an Odyssey infrared imaging system (Li‐Cor's Biosciences).