PM Exposure and Oxidative Stress Responses

Particulate matter (PM₁₀ PAH, PM_2.5 PAH, and PM_2.5 EI) was purchased from European Reference Materials (ERM-CZ100, ERM-CZ110; B-2440 European Commission, Geel, Belgium). PM_2.5 PAHs were also obtained by separation from the PM₁₀ certified reference material (i.e., ERM-CZ100) based on the modified sedimentation method (17 (link)). Liproxstatin-1 (SML1414), Deferiprone (379409), and Chloroquine (C6628) were purchased from Sigma-Aldrich Chemical Co. Inc. (St Louis, MO, USA). Necrosulfonamide (20844) and mito-TEMPO (16621) were purchased from Cayman Chemical Co. (Ann Arbor, MI, USA). z-VAD-fmk (cs-0015633) was purchased from ChemScene (Monmouth Junction, NJ, USA). N-acetyl-L-cysteine (A0905) was purchased from Tokyo Chemical Industry Co., Ltd. (Chiyodaku, Tokyo, Japan). The following antibodies were used: HO-1 (ADI-SPA-816; Enzo Biochem Inc., NY, USA), xCT (NB300-318; Novus Biologicals, CO, USA), transferrin receptor (13-6800; Invitrogen, MA, USA), ferritin (ab75973) and transferrin (ab1223; Abcam, Cambridge, UK), LC3B (L7543), and p62 (P0067; Sigma-Aldrich Chemical Co. Inc., MO, USA). The proteins and inhibitors, β-actin (sc-47778), p53 (sc-6243), cleaved Caspase-9 (sc-56073), Keap1 (sc-33569), Nrf2 (sc-722), ATG5 (sc-133158), and ATG7 (sc-33211) were obtained. SOD1 (sc-11407), SOD2 (sc-30080), catalase (sc-50508; Santa Cruz Biotechnology Inc., CA, USA), cleaved Caspase-3 (9661s), cleaved PARP (5625s), and GPX-4 (52455s; Cell Signaling Technology Co., MA, USA).