Anti atf4

24S-OHC [52 (link)] was dissolved in EtOH (Wako, Osaka, Japan). F12511 was the generous gift of Kowa (Aichi, Japan). Thapsigargin and CHX were purchased from Wako (Osaka, Japan). GSK2606414 and ISRIB were from Cayman Chemical (Ann Arbor, MI, USA). GCN2iB was from MedChemExpress (Monmouth Junction, NJ, USA). Thapsigargin, GSK2606414, CHX, and ISRIB were dissolved in dimethyl sulfoxide (DMSO; Wako). The following antibodies were from commercial sources: anti-PERK (Cat# 3192), anti-phospho-eIF2α (Cat# 3398), anti-eIF2α (Cat# 5324), and anti-GCN2 (Cat# 3302) were from Cell Signaling (Danvers, MA, USA); anti-β-actin (Cat# A5441) was from Sigma-Aldrich (St. Louis, MO, USA); anti-TIA1 (Cat# 12133-2-AP), anti-ATF4 (Cat# 10835-1-AP), and anti-TDP-43 (Cat# 12782-2-AP) were all from Proteintech (Chicago, IL, USA); anti-G3BP1 (Cat# 611126) was from BD Biosciences (Franklin Lakes, NJ, USA); anti-phospho-GCN2 (Cat# ab75836) was from Abcam (Cambridge, UK); and anti-puromycin (Cat# MABE343) was from Merck Millipore (Burlington, MA, USA); All other chemicals, of analytical grade, were obtained from Sigma-Aldrich or Wako.

All reagents were purchased from Sigma‐Aldrich (St. Louis, MO, USA), Wako (Osaka, Japan), or Takara (Kyoto, Japan), unless otherwise stated. Anti‐DYKDDDDK antibody (Wako), anti‐TMEM160 antibody (Invitrogen, Carlsbad, CA, USA), anti‐VDAC1 antibody (Calbiochem‐Novabiochem, San Diego, CA, USA), anti‐HSPA9 (Sigma‐Aldrich), anti‐TOMM22 (Tom22) antibody [4 (link)], anti‐HSPD1 antibody (Sigma‐Aldrich), anti‐glyceraldehyde‐3‐phosphate dehydrogenase (anti‐GAPDH) antibody (Novus Biologicals, Littleton, CO, USA), anti‐LONP1 antibody (Sigma‐Aldrich), and Total OXPHOS Rodent WB Antibody Cocktail (Abcam, Cambridge, UK) containing anti‐ATP5A, UQCRC2, and MTCO1 antibodies were used to immunostain the cells and/or in immunoblot analysis. The anti‐TOMM40 (Tom40) and anti‐TOMM20 antibodies were obtained previously [5 (link), 6 (link)]. Anti‐ATF4, ATF5, and DDIT3 antibodies were purchased from Proteintech (Rosemont, IL, USA).

Total ovary lysates were separated on SDS-PAGE. After electrophoresis, the protein was transferred to PVDF membrane. The membranes were blocked with 5% skimmed milk and incubated with anti-GRP78 (1:1000, #11587–1-AP, Proteintech, Hubei, China), anti-PARP1 (1:4000, #66520-1-lg, Proteintech), anti-PAR (1:1000, #4335-MC-100, Bio-techne, Minneapolis, USA), anti-mTOR (1:1000, #20657-1-AP, Proteintech), anti-p-mTOR (1:800, #sc-293133, Santa cruz, Dallas, Texas, USA), anti-KitL (1:1000, #bs-0545R, Biorbyt, Hubei, China), anti-p-IRE1α (1:800#, bs-16698R, Biorbyt), anti-IRE1α (1:1000, #27528-1-AP, Proteintech), anti-ATF4 (1:1000, #10835-1-AP, Proteintech), anti-p-PERK (1:800, #Orb336657, Biorbyt), anti-PERK (1:1000, #24390-1-AP, Proteintech), anti-HA-tag (1:1000, #30701ES60, YEASEN), anti-FLAG-tag (1:1000, #30501ES60, YEASEN) and anti-GAPDH (1:4000, #ab8245, Abcam, Cambridge, UK) antibodies overnight at 4 °C. After TBST washing, the membranes were treated with the corresponding peroxidase-conjugated secondary antibody, and then the signal was detected with an Enhanced Chemiluminescence Detection Kit (#32106, Thermo Scientific, Waltham, MA, USA) on Tanon 4500 gel imaging system (Shanghai, China).