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. Author manuscript; available in PMC: 2018 Apr 20.
Published in final edited form as: Cell Chem Biol. 2017 Apr 6;24(4):507–514.e4. doi: 10.1016/j.chembiol.2017.03.009

Figure 2. Val-boroPro activates caspases-3/7 in GSDMD knockout cells. See also Figs. S1-4.

Figure 2

(A) Immunoblotting reveals PARP cleavage in GSDMD−/−, but not in control or CASP−/−, THP-1 cells treated with Val-boroPro. In control cells, Val-boroPro induces the cleavage of GSDMD into the pyroptotic p30 fragment. Etoposide unexpectedly triggered the formation of a p43 fragment of GSDMD in control and CASP−/− cells. (B) Val-boroPro induces Annexin V positive/propidium iodide (PI) negative staining in GSDMD−/− THP-1 cells. Data are means ± SEM of 3 independent experiments. *** p < 0.001 compared to DMSO treated cells. (C–E) Effects of Val-boroPro, LPS plus nigericin, and etoposide on PARP and GSDMD in wild-type (C), GSDMD KO1 (D), and CASP1 KO1 (E) THP-1 cells as determined by immunoblotting. Nig, nigericin. (F) Val-boroPro induces cleavage of caspases-3 and -7 in GSDMD knockout cells. (G) Caspase-3/7 activity is elevated in GSMD KO THP-1 cells treated with Val-boroPro. Data are means ± SEM of 3 independent experiments. *** p < 0.001 compared to DMSO treated cells.