Goat anti mouse 115 035 003

For protein expression analysis, 293T cells were transfected with pcDNA3.1-gE, pcDNA3.1-IE63, pcDNA3.1-IE63-2A-gE, or pcDNA3.1-IE63-linker-gE. Forty-eight hours after transfection, cells and supernatants were obtained and analyzed using western blotting. After electrophoresis, proteins were transferred onto nitrocellulose membranes. Membranes were first incubated with a mouse-specific primary antibody against gE (ab272686; Abcam, UK) or rabbit polyclonal antibodies against IE63 (a gift from Dr. Paul R. Kinchington of University of Pittsburgh) for two hours and then with the secondary goat anti-mouse (115-035-003; Jackson ImmunoResearch Inc., West Grove, PA, USA) or goat anti-rabbit IgG-horseradish peroxidase (HRP) (111-035-045; Jackson ImmunoResearch Inc.) for 45 min. The nitrocellulose membrane was incubated with ECL luminescence reagent (Meilunbio, China, Cat. MA0186-1). Images were captured using an ECL system (Tanon, China).

HaCaT and PIG1 cells were lysed in RIPA lysis buffer on ice. Equal amounts of protein from each sample were separated using 10% SDS-PAGE and then transferred to polyvinylidene fluoride membranes (Merck Millipore, Billerica, MA, USA). In addition, 5% bovine serum albumin (BSA) (Yeasen, Shanghai, China) was used to block nonspecific binding for 1 h. The membranes were probed with anti-Keap1 (ab227828), anti-Nrf2 (ab62352), anti-HO-1 (ab13248), and anti-β-actin (ab8227) antibodies overnight at 4°C; these primary antibodies were obtained from Abcam (Cambridge, MA, USA). Membranes were then incubated with secondary antibodies, including goat anti-mouse (115-035-003; Jackson ImmunoResearch, PA, USA) and goat anti-rabbit (111-035-003, Jackson ImmunoResearch, PA, USA) antibodies, at room temperature for 1 h. Thereafter, the protein bands were examined using an ECL western blot detection system (Merck Millipore, Burlington, MA, USA). The antibodies used are listed in Supplementary Table 2.

Chloroquine (C6628) was obtained from Sigma-Aldrich. Oxaliplatin (OXA, S1224), and quinacrine (S5435) were purchased from Selleckchem. Torin 1 (2273-5) was ordered from BioVision. Anti-ATF4 (CST 11815S), anti-CHOP antibody (2895), anti-eIF2α antibody (9722), anti-p-eIF2α (Ser51) (9721S), anti-Histone H3 (9715S), anti-ATG5 antibody (12994), and anti-PERK (5683) antibodies were obtained from Cell Signaling Technology (CST). An anti-LC3 antibody (NB100-2220) was obtained from Novus Biologicals. An anti-GAPDH antibody (GTX100118) was ordered from GeneTex. Anti-TFE3 antibody (HPA023881), and anti-Flag (F3165) antibody were ordered from Sigma-Aldrich. Anti-TFEB antibody (303-673A) was ordered from Bethyl Laboratories. Anti-HMGB1 (ab18256), and anti-Calreticulin (ab92516) antibodies were ordered from Abcam. Goat anti-rabbit (111-035-003), and goat anti-mouse (115-035-003) secondary antibodies were products of Jackson ImmunoResearch. Alexa Fluor 594 goat anti-rabbit IgG (A-11012), Alexa Fluor 488 goat anti-rabbit IgG (A-11034), and Alexa Fluor 488 (A-11008) goat anti-rabbit secondary antibodies were ordered from Invitrogen.