Vdac2

Western blotting and quantitative PCR (RT‐qPCR) were performed following the previous report.
⁴³ (link) Following antibodies were used in Western blotting: Actin, FSP1, ACSL4, SLC7A11, VDAC2, and GPX4 rabbit polyclonal antibodies (1:1000; Abcam); VCAM‐1 and ICAM‐1 (1:500; Santa Cruz Biotechnology, Inc, Dallas, TX, USA). The primers used in RT‐qPCR are shown in Table S1.

The total lysates from the parental and VDAC1 knock-out HAP1 cells were obtained from 10⁶ cells in a lysis buffer (50 mM Tris-HCl pH 7.4, 150 mM NaCl, EDTA 1 mM, 1% TRITON X-100, protease inhibitors cocktail). Protein samples were separated using SDS-PAGE electrophoresis and transferred to a PDVF membrane (GE Healthcare, Chicago, IL, USA). The membranes were blocked in 5% BSA in PBS with 0.1% Tween-20 at room temperature for 1h and incubated overnight at 4 °C with the following primary antibodies: VDAC1 (1:1000, Abcam, Cambridge, UK), VDAC2 (1:200, Abcam), VDAC3 (1:100, Abcam), and β-Tubulin (1:2000, Cell Signaling, Danvers, MA, USA). After washing, membranes were incubated with IRDye-conjugated secondary antibodies (1:25,000, Li-Cor Biosciences, Lincoln, NE, USA). Signals were detected by the Odyssey CLx Imaging System (Li-Cor Biosciences) and analyzed using Image Studio Lite software (Li-Cor Biosciences). The same software was used for quantification.