N acetyl l cysteine a7250

NSC676914A (Additional file 1: Figure S1A) was synthesized by diazotization of 2-[(4-aminophenyl)sulfonyl]-1-hydrogen sulfate followed by coupling with m-toluidine. An unsulfated alcohol analog (Additional file 1: Figure S1B) was similarly prepared from 2-[(4-aminophenyl)sulfonyl]-1-ethanol. Both compounds were purified to >95% purity by reverse phase HPLC. DMSO stocks were used for all experiments. IKKβ inhibitor (IKK-2 Inhibitor IV [5-(p-Fluorophenyl)-2-ureido]thiophene-3-carboxamide, catalog #401484) (Additional file 1: Figure S1C) was purchased from EMD Biosciences (La Jolla, CA). LPA (857130C) was purchased from Avanti Polar Lipids (Alabaster, AL). LPA stocks were made in PBS containing 1% fatty acid-free bovine serum albumin. TPA (4174) was purchased from Cell Signaling Technology, Inc (Danvers, MA). Recombinant Human TNFα (300-01A) was purchased from Peprotech (Rocky Hill, NJ). Z-VAD-FMK (2163) and Necrostatin-1 (2324) were purchased from Tocris Bioscience (Ellisville, MO). Puromycin, XTT, PMS, LPS (L5293) and N-Acetyl-L-cysteine (A7250) were purchased from Sigma-Aldrich (St. Louis, MO).

The ethyl alcohol (pure (E7023) and FastStart Essential DNA Green Master (06402712001)) were obtained from Sigma-Aldrich (Darmstadt, Germany). The 16% formaldehyde (methanol-free) was obtained from Thermo Fisher Scientific (28906, Seoul, Republic of Korea). The μ-Dish 35mm image dishes with polymer coverslip bottoms were obtained from ibidi GmbH (Gräfelfing, Germany). N-acetyl-L-cysteine (A7250) and MitoTEMPO (SML0737) were obtained from Sigma-Aldrich (Darmstadt, Germany).
The antibodies against RAP1 were obtained from Bethyl laboratories, Inc. (A300-306A-9, MA, USA); the anti-TPP1 (ab195234), anti-TRF1 (ab10579), anti-p16 (ab211542), and anti-53BP1 (ab175933) were from Abcam (Cambridge, UK); the anti-POT1 (NB500-176) and anti-TRF2 (NB100-56506) were from Novus Biologicals (Littleton, CO, USA); the anti-TIN2 (PA5-67076) was from Invitrogen (Waltham, MA, USA); the anti-p53 (P6874), p16 (ab211542), and anti-actin (A5441) were from Sigma-Aldrich (Darmstadt, Germany); and the anti-P21 (2947), PINK1 (6946), Parkin (4211), autophagy sampler kit (4445), SirT3 (2627), SirT5 (8779), and Tom20 (42406) were obtained from Cell Signaling Technology (Danvers, MA, USA). Ginsenoside F1 was prepared as described in our previous report [34 (link)].