Fig. 7: USP18-STAT2 regulates GSDMD and GSDME-dependent pyroptosis upon IFN stimulation.
From: Expansion of interferon inducible gene pool via USP18 inhibition promotes cancer cell pyroptosis
a Western blot of WT, GSDMD−/−, GSDME−/−, USP18−/−, USP18−/−GSDMD−/− and USP18−/−GSDME−/− THP-1 cells treated with or without IFNα (1000 U/ml) for 48 h. b Analysis of ICD markers in (a). c Percentage of live cells in (a) analyzed by FACS. d Western blot of WT or USP18−/− cells expressing Control sgRNA or Caspase-1 sgRNA after treatment with IFNα (1000 U/ml) for 48 h. e Analysis of IL-1β secretions of culture supernatant in (d). f Percentage of live cells in (d) analyzed by FACS. g Western blot of DMSO, MCC950 or ODN-A151 treated WT or USP18−/− THP-1 cells further treated with or without IFNα (1000 U/ml) for 48 h. h Analysis of IL-1β secretions of culture supernatant in (g). i Percentage of live cells in (g) analyzed by FACS. j Western blot of USP18−/− THP-1 cells expressing Mock, USP18 or USP18C64S treated with or without IFNα for 48 h. k Percentage of live cells in (j) analyzed by FACS. l Analysis of ICD markers in (j). m qRT-PCR of IFIT1, PLK2 and CCL4L2 expression in USP18−/− THP-1 cells expressing Mock, USP18 or USP18C64S treated with or without IFNα (1000U/ml) for 6 h. n Percentage of live WT, USP18+/− and USP18−/− 2fTGH cells after 48 h of treatment with or without IFNα (1000 U/ml). o Analysis of ICD markers in (n). p qRT-PCR of IFIT1 and PLK2 expression of STAT2 or STAT2 R148W expressing STAT2−/− (U6A) cells that were treated with or without IFNα (1000 U/ml) as indicated. q Percentage of live cells of STAT2 or STAT2 R148W expressing STAT2−/− (U6A) cells that were treated with or without IFNα (1000 U/ml) for 48 h. r Western blot of cell lysates in (q). s Analysis of IL-1β secretions of culture supernatant in (q). t Analysis of ICD markers in (q). n = 2 for Fig. 7p and n = 3 independent samples for (b, c, e, f, h, I, k–o, q, s–t) were analyzed. All data represent mean ± s.d, except where indicated. n.s. = not statistically significant. All p-value in Fig. 7 was determined by one-way ANOVA test. Source data are provided as a Source Data file.
