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. 2019 Sep 30;17(9):e3000402. doi: 10.1371/journal.pbio.3000402

Fig 3. Tumor cell death in response to CRT down-regulation is executioner caspase independent.

Fig 3

(A) Western blot analysis of PARP full-length and apoptosis specific cleaved protein fragments (cPARP) in an indicated types of malignant cells following CRT knockdown. (B) Western blot analysis of full-length and cleaved PARP fragment following treatment of cells with apoptosis inducing agent, SS. (C) Western blot analysis of necrosis-specific PARP cleavage fragments using C2-10 anti-PARP mAb in the indicated solid-tissue malignancies transduced with shCRT and shCont. Western blot analysis of the cleaved Caspase-3 (D) and Caspase-7 (E) proteins in the cells following CRT knockdown or SS treatment. (F) Immuno blot of total BAX after IP with antiactive-BAX antibody; 10% of IP input was used as a loading control for BAX. BAX, BCL2-associated X protein; cPARP, cleaved poly ADP ribose polymerase; CRT, calreticulin; GAPDH, Glyceraldehyde 3-phosphate dehydrogenase; HCT, Human colorectal carcinoma cell line; IB, immunoblot; IP, immunoprecipitation; mAb, monocloncal antibody; PARP, poly ADP ribose polymerase; RCC, Human clear cell renal cell carcinoma cell line; shCont, short hairpin RNA targeting Control; shCRT, short hairpin RNA targeting Calreticulin; SS, Staurosporin.