Nac a7250

Antiretroviral drugs TDF, FTC (Gilead Sciences, Foster City, CA, USA), and DTG (ViiV Healthcare, Research Triangle Park, NC, USA) were used. NAC (A7250) and (2-(2,2,6,6-Tetramethylpiperidin-1-oxyl-4-ylamino)-2-oxoethyl)triphenylphosphonium chloride (mitoTEMPO; SML0737) were purchased from Sigma-Aldrich, St. Louis, MO, USA, and 4-hydroxy-2,2,6,6-tetramethylpiperidin-1-oxyl (TEMPOL; sc-200825) was purchased from Santa Cruz Biotechnology, Dallas, TX, USA. Antibody resources: beclin 1 (BECN1; sc-11427) was purchased from Santa Cruz Biotechnology, Dallas, TX, USA. Lysosome associated membrane protein 2 (LAMP2; NB300-591), microtubule-associated protein 1 light chain 3 beta (MAP1LC3B; NB100-2220), and integrin subunit alpha M (ITGAM; NB110-89474) were purchased from Novus Biological Company, Centennial, CO, USA. Cathepsin D (CTSD; ab75852) and caspase 3 (CASP3; ab13585) were purchased from Abcam, Cambridge, MA, USA. Galectin 3 (GAL3) (A3A12) was purchased from Invitrogen. Sequestosome 1 (SQSTM1; PM045) was purchased from MBL International, Woburn, MA, USA, and allograft inflammatory factor 1 (AIF1; 019-19741) was purchased from Wako Pure Chemicals Industries, Chuo-ku, Osaka, Japan. Goat anti-rabbit (sc-2004) and goat anti-mouse (sc-2005) were purchased from Santa Cruz Biotechnology, Dallas, TX, USA.

Sodium Iodate (NaIO₃; 71702) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Retinal Pigment Epithelial Cell Growth Medium (RtEGM^TM; #00195407) with supplements including 2% FBS, 2% L-glutamine, 0.5% bFGF, 0.1% GA-1000 was purchased from LONZA (Walkersville, MD, USA). Dulbecco’s modified Eagle’s medium (DMEM; 12800-017), fetal bovine serum (FBS; 26140-079), penicillin/streptomycin (10378-016), and 0.25% trypsin (25200-072) and other cell culture reagents were purchased from Gibco (Gaithersburg, MD, USA). Primary antibodies including phosphor-Akt (ser473; #4058, Thr308; #9275), total Akt (#9272), phospho-ERK (#4370), ERK (#4695), phosphor-JNK (#9251), JNK (#9252), phosphor-p38 (#4511), p38 (#9212), phosphor-IκBα (#9246), and IκBα (#9242) (Cell Signaling Technology, Inc., Danvers, MA, USA), and β-actin polyclonal antibody (SC-47778) (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) were used for western blotting analysis. Human pentraxin 3 (PTX3; DY1826) ELISA kit was purchased from R&D System, Inc. (Minneapolis, MN, USA). U0126 (BML-EI282), LY194002 (BML-ST420), SP600125 (BML-EI305), SB203580 (BML-EI286), BAY 11-7082 (BML-EI278) (Enzo Life Sciences, Farmingdale, NY, USA), and NAC (A7250) (Sigma-Aldrich, St. Louis, MO, USA) were used for inhibitor reagents.

All chemical and biochemical reagents were purchased from the following sources: DMEM from Nacalai Tesque (Kyoto, Japan), FBS from Thermo Fisher Scientific (Rockford, IL, USA), and NAC (A7250) from Sigma-Aldrich (Burlington, MA, USA). The total ROS detection kit was obtained from Dojindo (Kumamoto, Japan). Antibodies against ERK, p-ERK, Akt, p-Akt, LC3B, p62, and β-actin were obtained from Cell Signaling Technology (Beverly, MA, USA). Antibodies against LC3 and p62 were obtained from MBL (Nagoya, Japan).