Ab81212

HCT116 human colon carcinoma cells were obtained from the American Type Culture Collection (CCL-247). Artesunate is purchased from Sigma Aldrich (A3731, St. Louis, MO, USA). Antibodies for western blotting are purchased from Cell Signaling Technology (Danvers, MA, USA) and Abcam (Cambridge, UK): BCS1 (Abcam; ab102808), UBP1 (Abcam; ab30965), PDXK.1 (Abcam; ab119051), AOC2 (Abcam; ab83734), CDK11 (CST; 5524), Actin (CST; 3700), ACSL5 (Abcam; ab57210), HADH (Abcam; ab54477), FASN (CST; 3180), NDA (Abcam; ab81212), COX (Abcam; ab110267), Cyt-c (CST; 11940), TIM50 (Abcam; ab23938), Bax (CST; 5023), Bcl-2 (CST; 15071), AIF (Abcam; ab32516), Cleaved PARP (CST; 5625), Caspase 9 (CST; 9502), NF-κB p105 (CST; 12540), PP2Aα (CST; 2041), PP2Aβ (CST; 4953), USP15 (Abcam; ab56900), IκB (CST; 9247), p-p65 (CST; 3033), p65 (CST; 8242).

Rabbit anti-CDT-1 (Bethyl A300-786A, 1:1,000), rabbit anti-FAF1 (Cell Signaling #4932S, 1:1,000), mouse anti-β-actin (Abcam Ab6276, 1:10,000), mouse anti-p97 (Abcam Ab11433, 1:1,000), mouse anti-ATM (Sigma-Aldrich A1106, 1:2,000), rabbit anti-phospho ATM ser1941 (Abcam Ab81212, 1:5,000), rabbit anti-ATR (Bethyl A300-A137A, 1:1,000), rabbit anti-phospho ATR ser428 (Millipore ABE389, 1:1,000), mouse anti-CHK-1 (Cell Signaling 2360, 1:1,000), rabbit anti-phospho CHK-1 ser345 (Cell Signaling 2348, 1:1,000), rabbit anti-CHK2 (Cell Signaling 2662, 1:1,000), rabbit anti-phospho CHK2 Thr68 (Cell Signaling 2661), rabbit anti-phospho H2AX Ser139 (Novus Biologicals NB100-2280, 1:1,000), rat anti-HA (Roche, clone 3F10, 1:5,000) and mouse anti-PCNA (Abcam Ab29, 1:1,000). Anti-rabbit and anti-mouse horse radish peroxidase-conjugated secondary antibodies were purchased from Sigma-Aldrich (A9169 and A9044, respectively, 1:10,000). Membranes were activated using Thermo Scientific SuperSignal West Pico and Dura substrates and developed accordingly using standard protocol.

Western blotting was conducted as in the previous studies [19 (link), 20 (link)]. Primary antibodies used herein include those targeting ND6 (Abcam, ab81212), ND4 (Abcam, ab219822), ATP6 (Proteintech, 55313-1-AP), ND1 (Abcam, ab74257), CYB (Proteintech, 55090-1AP), CO2 (Abcam, ab110258), and VDAC (Proteintech, 10866-1-AP). As a secondary detection antibody, we employed the peroxidase AffiniPure goat anti-mouse IgG and goat anti-rabbit IgG (Jackson) constructs, and an enhanced chemiluminescence (ECL) system (Millipore) was used for band detection, after which densitometric calculations were performed.