Hpa001401

Western blot was performed as previously described [11 ]. Equal amount of protein was boiled in LDS sample buffer containing reducing agent while for WB-CETSA 10 µl of protein lysate from each sample was used. Primary antibodies used were horseradish peroxidase (HRP)-conjugated antibody against p53 (DO-1) diluted 1:5000 (sc-126 HRP, Santa Cruz Biotechnology, USA) and GAPDH diluted 1:30,000 (sc-47724 HRP, Santa Cruz Biotechnology, USA). Primary antibodies without any conjugation were against ASNS diluted 1:1000 (sc-365809, Santa Cruz Biotechnology, USA), SOD1 diluted 1:20,000 (HPA001401, Sigma), xCT/SLC7A11 (D2M7A) diluted 1:1000 (12691S, Cell Signaling, USA), and TrxR1 diluted 1:1000 (Santa Cruz Biotechnology, USA).

Mouse monoclonal antibody 9E10 anti-HCV NS5A was kindly provided by Charles Rice^{38 (link)} and diluted 1:1000 in TCID₅₀ assays. Commercial antibodies used in Western blotting, along with their respective dilutions, are listed as follow. Mouse monoclonal antibody anti-HCV NS3 was purchased from Abcam (ab65407, 1:3,000). Mouse monoclonal antibody against GAPDH, rabbit polyclonal antibody against CRBN, and rabbit polyclonal antibody against SOD1 were respectively purchased from GeneTex (GTX28245, 1:5,000), Novus Biologicals (NBP1-91810, 1:500), and Sigma-Aldrich (HPA00140-1; 1:500). Horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG and anti-rabbit IgG antibodies were obtained from Bio-Rad Laboratories (170-6516 and 170-6515, respectively, 1:3,000). IRDye® 800CW-conjugated goat anti-rabbit antibody was purchased from LI-COR (926-32211, 1:10,000).