Camptothecin

To measure proliferation, 150µL of cells at 0.1 × 10⁶ cells/mL were plated in 96-well plates in triplicate for five days. Each day, cells were counted in triplicate using a cell counter (Beckman Coulter). For drug response assays, 2mL of cells were diluted to 0.5 × 10⁵ cells/mL in 12-well plates in their culture medium and treated with the corresponding agent, camptothecin (Tocris Bioscience), doxorubicin (Sigma-Aldrich), prednisolone (Sigma-Aldrich) to promote DNA damage-induced apoptosis, then harvested to assay for apoptosis. DMSO (vehicle) was used as control for prednisolone treatment, while other drugs were diluted in water. Cell apoptosis was measured using the Alexa Fluor® 488 Annexin V/Dead Cell Apoptosis Kit (Thermo Fisher Scientific) according to the manufacturer's instructions. Stained cells were analyzed by flow cytometry using the BD FACSCanto^TM II cell analyser and the BD FACSDiva software (BD Biosciences) according to manufacturer's guidelines.

Sorafenib, Camptothecin, and Dimethyl fumarate (DMF, Nrf2 inducer) were purchased from (Tocris Biosciences). Ferrostatin-1 and ZVAD-FMK were obtained from Sigma-Aldrich (Shanghai, China). Dimethyl sulfoxide (DMSO) and 3-(4,5-dimethyl thiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) were provided from (Sigma-Aldrich, USA, and Serva^® Germany respectively). Dulbecco's Modified Eagle's Medium (DMEM), penicillin, streptomycin, fetal bovine serum, and phosphate buffer saline (PBS) were obtained from (Gibco-USA). RIPA buffer (50 mM Tris–HCl (pH 7.4), 150 mM NaCl, 5 mM EDTA, 1% Triton X-100, 0.5% Sodium deoxycholate, and 0.1% SDS), HEPES buffer (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) were purchased from (Biowest, India). Antibodies against Nrf2, p62, SLC7A11, MT-1G, and ABCG2 were provided by (Abcam, UK).

Lymphoprep™ gradient was purchased from Progen (Heidelberg, Germany). RPMI 1640 medium, fetal calf serum, trypsin 10x (25 mg/mL), trypsin-EDTA 10x (5 mg/mL trypsin and 2.2 mg/mL EDTA), and phosphate buffered saline (PBS, without Ca and Mg) were from PAA Laboratories Gmbh (Coelbe, Germany). L-Glutamine, penicillin, and streptomycin were from Invitrogen (Karlsruhe, Germany). Camptothecin was from Tocris (Eching, Germany); Triton-X 100, milk powder, and N,N,N′,N′-tetramethyl-1-,2-diaminomethane (Temed) were from Carl Roth (Karlsruhe, Germany); DMSO, lipopolysaccharide (LPS), phorbol, 12-myristate 13-acetate, CFSE, ammonium persulfate, bovine serum albumin, ethanol absolute, hydrochloric acid (37%), leupeptin hemisulfate, p-coumaric acid, pepstatin A, Ponceau S, trypan blue, Tween 20, and ionomycin were from Sigma-Aldrich (Taufkirchen, Germany).
Antibodies against p-38 (Thr180/Tyr182), p-ERK1/2 (Thr202/Tyr204), p-JNK, and p-c-Jun (Ser73) and the horseradish peroxidase- (HRP-) labelled secondary antibodies, anti-mouse and anti-rabbit, were from Cell Signaling Technology (Boston, USA); anti-human COX-2 was from R&D Systems (Wiesbaden, Germany); anti-human COX-1 and anti-human 5-LOX (mouse monoclonal, clone 33) were from Santa Cruz Biotechnology (Heidelberg, Germany); and mAb against β-actin was from Sigma-Aldrich (Taufkirchen, Germany).