Ab102808

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).

Heart tissues were processed for western blotting as described previously. Samples were disrupted using a ice-cold radioimmunoprecipitation assay buffer containing 10 mM Tris–Cl at pH 8.0, 1 mM ethylenediamine tetraacetic acid, 1% Triton X-100, sodium deoxycholate at 0.1%, and sodium dodecyl sulfate, also at 0.1%, enriched with 150 mM NaCl, 1 mM phenylmethylsulfonyl fluoride, and a mixture of protease inhibitors: aprotinin, leupeptin, and pepstatin at 0.02 mg/mL each. Following sonication, these lysates underwent clarification by centrifugation. Protein quantification was conducted utilizing the Bradford assay, and aliquots containing 50 to 100 mg of protein per lane were subjected to separation via sodium dodecyl sulfate–polyacrylamide gel electrophoresis. Postseparation, these proteins were electroblotted onto a polyvinylidene difluoride membrane and probed with specific antibodies, namely BCS1L (1:500, no. ab102808, Abcam) and FBXO7 (1:1000, no. ABN1038, Merck).