Fig. 5: Apoptotic response to X-rays, WS-12 or their combination in RWPE-1 based models of premalignant and malignant prostate lesions. | Cell Death & Disease

Fig. 5: Apoptotic response to X-rays, WS-12 or their combination in RWPE-1 based models of premalignant and malignant prostate lesions.

From: Calcium cytotoxicity sensitizes prostate cancer cells to standard-of-care treatments for locally advanced tumors

Fig. 5

a Diagram of the experimental design. Genetically engineered RWPE-1 cells were treated as indicated (BioRender.com). b Representative flow cytometry analysis of cell death in ERG + and ERG + /PTEN-deficient RWPE-1 M8 cells treated with WS-12 (1 μM, 12 h), X-rays (10 Gy) or the combination of both. Untreated cells were used as control. c Cell death quantification of RWPE-1 M8 cells treated as described in b is reported as percentage of total cells. d Western blotting analysis of samples described in b shows CaMKIIα activation (phosphorylation of Thr286) following WS-12 treatment of RWPE-1 M8 and the classical molecular signature of apoptotic cell death (Caspase-3 and PARP cleavage) in RWPE-1 M8-based premalignant (ERG-shCTR) and malignant (ERG + shPTEN) models of human prostatic disease treated with X-rays plus WS-12. e Representative ERG+ and ERG+/PTEN-deficient RWPE-1 M8 3D prostopheres treated with WS-12 (1 μM, 12 h), X-rays (10 Gy) or the combination of both. Untreated spheroids were used as control. Scale bar, 50 μm. f Western blotting analysis of treated prostopheres described in e showing classical molecular hallmarks of apoptotic cell death (Caspase-3 and PARP cleavage). Error bars, mean ± SD. Experiments were performed in quadruplicate; data were analyzed using a two-way ANOVA test. **P ≤ 0.01.

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