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. 2020 Feb 24;11(2):149. doi: 10.1038/s41419-020-2335-1

Fig. 5. STAT5 protects cells from pimozide-mediated cell death.

Fig. 5

a Cells were transfected with siRNA for STAT5A, STAT5B or both for 72 h. Lysates were analyzed STAT5 western blot. Knockdown of STAT5A or STAT5B reduced STAT5 protein levels, with greater reduction observed in cells transfected with STAT5B. b Cells were transfected with siRNA for STAT5A, STAT5B or both for 48 h, and subsequently subjected to flow cytometry following staining with propidium iodide. Knockdown of STAT5 protein did not affect cell cycle progression of OS cells. c Cells transfected with siRNA to STAT5A, STAT5B or both were treated with 10 µM pimozide. Cell proliferation measured by hexosaminidase assay demonstrated that knockdown of STAT5 reduced cell proliferation. d Cells were transfected with siRNA for STAT5A, STAT5B or both for 48 h and then treated with 10 µM pimozide for 24 h. Lysates were analyzed for STAT5 and Caspase 3 levels. Knockdown of STAT5A or STAT5B further enhanced pimozide mediated caspase 3 cleavage. e Cells were transfected with siRNA to STAT5A, STAT5B or both and then treated with pimozide were analyzed by flow cytometry following AnnexinV/PI staining. Pimozide treatment significantly induced cells to undergo necrosis and late apoptosis. However, knockdown of STAT5B reduced pimozide mediated necrosis resulting in further increase in apoptosis.