Anti phospho p38

The relative amount of various proteins was determined by western blot analysis according to the protocol as described.²⁹ The antibodies used for immunoblot analysis were purchased from Cell Signaling Technologies (CST), Santa Cruz Biotechnology (SCBT) or R&D Biosystems (R&D): anti‐Ubiquitin (SCBT sc8017), anti‐Beclin‐1 (CST 3495), anti‐light chain (LC)‐3B (CST 2775), anti‐GAPDH (CST 2118), anti‐phospho‐p65 (CST 3033), anti‐p65 (CST 8242), anti‐phospho‐p38 (CST 9211), anti‐p38 (CST 9212), anti‐phospho‐AMPK (CST 2535), anti‐AMPK (CST 2532), anti‐TAK1 (CST 4505), anti‐Nrf2 (R&D Systems, MAB3925), and anti‐KEAP1 (CST 8647). HRP conjugated secondary antibodies were procured from Cell Signaling Technology. Band intensities were quantified with Image J software.

DOX was purchased as Adriablastine (50 mg doxorubicin hydrochloride, Pharmacia & Upjohn, Milan, Italy). Chrysin, GSH, Ellman’s reagent [5,5-dithio-bis (2-nitrobenzoic acid); DTNB], bovine serum albumin, dimethylsulphoxide (DMSO) and thiobarbituric acid were purchased from Sigma Chemical Co. (St Louis, MO, USA). Anti total p38, anti phospho p38, anti total JNK and anti phospho JNK antibodies were purchased from R & D systems (Minneapolis, USA). Anti total AKT and anti phospho AKT antibodies were purchased from Biovision Co. (San Francisco, USA). Anti GAPDH antibody was purchased from Santa cruz biotechnology (Santa cruz, Texas, USA). Anti NF-κB p65, VEGF, p53, cytochrome c, caspase 3 antibodies were purchased from Thermo Fisher Scientific Co. (Waltham, MA, USA). All other chemicals were of the highest purity grade commercially available.