Cleaved parp asp214 d64e10

The p53 (DO-1), PUMAα (B-6), and vinculin (7F9) antibodies were from Santa Cruz Biotechnology (Dallas, TX, USA). The antibodies against DR5 (D4E9), cleaved caspase 3 (D175) and cleaved PARP (Asp214) (D64E10) were from Cell Signaling Technology (Danvers, MA, USA). The HA tag (clone 3F10) and β-actin (clone AC-15) antibodies were from Roche Applied Science (Penzberg, Germany) and Sigma-Aldrich (St. Louis, MO, USA), respectively. ECRG2 antibody was generated in our laboratory as previously described^{13 (link)}. The peroxidase-conjugated horse anti-mouse, goat anti-rat, and goat anti-rabbit antibodies were from Vector Laboratories (Burlingame, CA, USA). The cells were transfected using PolyJet or LipoJet reagents (SignaGen Laboratories, Rockville, MD, USA). The plasmid subcloning was performed using the restriction endonucleases from New England BioLabs (Ipswich, MA, USA). Etoposide, doxycycline, thapsigargin, and melphalan were from Sigma-Aldrich (St. Louis, MO, USA), and sulindac sulfide was provided by Merck (Rahway, NJ, USA). The cells growing in logarithmic phase were exposed to UVC as described previously^{50 (link)}, except XL-1500 UV crosslinker (Spectronics, Westbury, NY, USA) was used as a source of UV radiation.

Cells were harvested after 30 h of treatment with Tg, unless otherwise stated. Preparation of whole-cell lysates, SDS-PAGE, and immunoblot analysis were performed as described previously [33 (link)]. The following antibodies were used: alpha-tubulin (Abcam, ab7291), ATG5 (Cell Signaling Technology 2630), ATF4 (Cell Signaling Technology 11815), Bip/Grp78 (Cell Signaling Technology 3177), Caspase-3 (8G10) (Cell Signaling Technology 9665), Caspase-8 (1C12) (Cell Signaling Technology 9746), CHOP (Cell Signaling Technology 2895), Cleaved PARP (Asp214) (D64E10) (Cell Signaling Technology 5625), DR5 (Cell Signaling Technology 8074), FADD (G-4) (Santa Cruz sc-271748), FIP200 (Proteintech 17250–1-AP, GABARAP (Proteintech PM037), GABARAPL1 (Abcam ab86497), GABARAPL2 (Proteintech PM038), p-JNK (Cell Signaling Technology 9251), JNK (Cell Signaling Technology 9252), LC3B (Cell Signaling Technology 2775), PERK (Cell Signaling Technology 5683), XBP1s (BioLegend 647502).

Protein extraction and Western blot analysis were performed as previously described [52 (link)]. The following primary antibodies were used: FoxM1 (D12D5), Cleaved PARP (Asp214) (D64E10), RhoA (67B9), Rac1/2/3, Cdc42 (11A11) (Cell Signaling Technology, Inc., Danvers, MA, USA), T7-tag monoclonal antibody (Merck KGaA, Darmstadt, Germany), GAPDH (Trevigen, Inc., Gaithersburg, MD, USA). Donkey anti-rabbit IgG-HRP (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) and anti-mouse IgG-HRP (Promega Corp., Madison, WI, USA) were used as a secondary antibody.