Full length parp

Whole cell lysates were prepared from cell lines with RIPA lysis buffer (100 mM Tris pH 7.5, 150 mM sodium chloride, 0.1% deoxycholate, 0.1% SDS, 50 mM NaF, Protease inhibitor cocktail (Roche), 2 mM PMSF, 2 mM sodium orthovanadate) combined with scraping and the lysates cleared by centrifugation. Standard Western blotting procedures were performed. The following primary antibodies were used for immunoblotting at a dilution of 1:1000: ZEB1 (3396, Cell Signaling Technology, Danvers, MA, USA), Full-length PARP (9532, Cell Signaling Technology), Hsp90 (sc-13119, Santa Cruz, Dallas, TX, USA) and β-actin (MABT825, MilliporeSigma). Treatments with rhTGFBI (R&D Systems, Minneapolis, MN, USA) or vehicle control (PBS) were performed at the specified doses for 24 h prior to harvesting lysates. All blots or gels derive from the same experiment and were processed in parallel. See Supplementary Information for unedited blots (Supplementary Fig. 8a–c).

Cell culture media and chemical reagents including Dulbecco’s modified Eagle’s medium (DMEM), minimum essential medium (MEM), Opti-MEM, Roswell Park Memorial Institute (RPMI) 1640, sodium pyruvate, and antibiotics were obtained commercially (Life Technologies, Grand Island, NY, USA). Fetal bovine serum (FBS) was used according to the manufacturer’s instructions (HyClone, Logan, UT, USA). Commercial antibodies were used to detect Src, phosphorylated-Src (Tyr416), STAT3, phosphorylated-STAT3 (Tyr705), JAK2, cleaved caspase-3, cleaved PARP, full-length PARP (Cell Signaling Technology, Beverly, MA, USA), Bax, Bcl-2, p53, survivin, and β-actin (Santa Cruz Biotechnology, Dallas, TX, USA). Antibody detection was done using goat anti-rabbit and anti-mouse antibodies coupled to horseradish peroxidase (HRP; Santa Cruz Biotechnology).