Figure 1.
VbP induces pyroptosis in pHCE cells. The pHCE cells were treated with DMSO or the DPP8/9 inhibitor VbP (10 µM) for 24 hours. Supernatants were evaluated for the release of LDH (A) and bioactive cytokines (B, C). Cell lysates and concentrated supernatants were assessed by immunoblotting (D). (E) Cells were treated with VbP (10 µM) and imaged by time-lapse microscopy. Representative images of propidium iodine (PI; red), and nuclei (Hoechst; blue) merged with brightfield (left panel); and zoom-in of pyroptotic cell, arrow heads reveal pyroptotic “blebs” (right panel). Scale bars 100 µm. Data are mean ± SEM from n = 3 donors A–C. ***P < 0.001, **P < 0.01 by unpaired t-test; WCL = whole cell lysate, sup = supernatant, GSDMDFL = full length GSDMD, GSDMDNT = N-terminal GSDMD p30 fragment resulting from caspase-1 cleavage.