Fig. 3.

Pyroptosis and caspase-3 activation are compromised in GSDME-T6E-expressing CEM-C7 cells. a–d Propidium iodide (PI) uptake (a, c), and active caspase-3 staining (b, d) in CEM-C7 cells (wild-type (WT), GSDME-knockout (KO), or GSDME-KO reconstituted with WT GSDME-EGFP or GSDME-T6E-EGFP) treated with triamcinolone acetonide (TA) (a, b) or ultraviolet (UV) (c, d) as measured on the IncuCyte over time. e, f Immunoblots of GSDME, active caspase-3 p17/p19 and high-mobility group box 1 (HMGB1) in combined cell lysates plus media, or HMGB1 released in culture media (media) of WT (WT), GSDME-KO (KO), or GSDME-KO reconstituted with WT GSDME-EGFP (KO + WT) or GSDME-T6E-EGFP (KO+T6E) CEM-C7 cells treated with TA (e) or UV (f). *Indicates GSDME-EGFP degradation product. Results are representative of at least three independent experiments performed in duplicate or triplicate. Error bars represent S.D. Student’s t-test, *p < 0.05, **p < 0.005, ***p < 0.0005