Lps hy d1056

The human esophageal cancer EC109, KYSE510 cell lines were obtained from the National Infrastructure of Cell Line Resource, cultured in RPMI-1640 medium (Gibco, Grand Island, NY, USA) with 10% fetal bovine serum (FBS) (Gibco), 100 units/mL penicillin (Invitrogen, Carlsbad, CA, USA) and 100 mg/mL streptomycin (Invitrogen). The cell lines were characterized by Genetic Testing Biotechnology Corporation (Suzhou, China) using short tandem repeat markers within the last three years and they were not contaminated by mycoplasma detected by Myco-Lumi Luminescent Mycoplasma Detection Kit (Beyotime, Shanghai, China). SFE was separated and purified from radish seeds (Beijing Tongrentang Co., LTD, Beijing, China) as reported previously [4 (link),5 (link)], dissolved in DMSO (Beijing Chemical Factory, Beijing, China). LPS (HY-D1056) and PDTC (HY-18738) was obtained from MedChem Express (Monmouth Junction, New Jersey, USA).

GSK2606414 (S7307) was ordered from Selleck (Pittsburgh, PA, USA), and LPS (HY-D1056) was ordered from MedChemExpress (Monmouth Junction, NJ, USA). The antibodies, including anti-LC3 (12741), anti-LC3-Ⅱ (3868), anti-P62 (5114), anti-ATG5 (12994), anti-ATG12 (4180), anti-Beclin-1 (3495), anti-p-PERK (Thr980, 3179), anti-p-eIF2α (Ser51, 3398), anti-P65 (8242), anti-p-P65 (Ser536, 3033), anti-Lamin B1 (13435), anti-LAMP1 (9091), anti-IκBα (4814), anti-p-IκBα (Ser32, 2859), anti-mTOR (2983T), anti-p-mTOR (ser2448, 5536), anti-ULK1 (6439), anti-p-ULK1 (ser555, 5869), anti-p-ULK1 (ser757, 14202), anti-AKT (2920), and anti-p-AKT (Ser473, 4060) were ordered from Cell Signaling Technology (Danvers, MA, USA); anti-TOM20 (11802-1-AP), anti-GM130 (11308-1-AP), anti-Bip (11587-1-AP), anti-ATF6 (24169-1-AP), anti-PERK (24390-1-AP), anti-eIF2α (11170-1-AP), anti-ATF4 (10835-1-AP), anti-XBP1 (25997-1-AP), anti-β-actin (20536-1-AP), and anti-GFP (50430-2-AP) were ordered from Proteintech (Rosemont, IL, USA); anti-Calnexin (C4731) was ordered from Sigma-Aldrich (St. Louis, MO, USA); and anti-TIA-1 (sc-166247) was ordered from SCBT (Dallas, TX, USA).