Horseradish peroxidase labeled antibodies

B-MD-C1 (wt) was purchased from ATCC, DOX (Adriamycin, ADR) was purchased from Hisun pharmaceuticals, Zhejiang, China and dissolved in sterile saline solution (2 mg/ml) and stored at -20 °C before use. Fetal bovine serum (FBS) and RPMI 1640 medium were purchased from Gibco, USA; Lipofectamine™2000 was purchased from Life Technologies, USA; antibodies against CRT, ERp57, caspase-8 and cleaved-caspase-8 were purchased from Abcam, USA; antibodies against p-PERK, eIF2α and p-eIF2α were purchased from Cell Signaling Technology, USA; β-actin and horseradish peroxidase-labeled antibodies were obtained from Santa Cruz, USA. MTT assay kit was obtained from Amresco, USA.

Total protein lysates or membrane-enriched protein extracts were prepared, separated by SDS-PAGE, and transferred onto a polyvinylidene difluoride membrane for Western blotting as described previously [54 (link)]. Primary antibodies used are given in Supplementary Table S6. Secondary, horseradish peroxidase-labeled antibodies from Santa Cruz Biotechnology were used in working dilutions of 1:10 000.

Total protein lysates of membrane-enriched extracts were prepared, separated by SDS-PAGE, and transferred onto a polyvinylidene difluoride membrane for western blotting as described previously (Heffeter et al. 2007 (link)). Primary antibodies used are given in Supplementary Table 2. Secondary, horseradish peroxidase-labeled antibodies from Santa Cruz Biotechnology were used in working dilutions of 1:10,000. Since reference proteins were affected by chronic arsenic treatment and could not be used for a reliable normalization, total protein load via Coomassie staining as outlined by (Eaton et al. 2013 (link)) was used instead.