Extended Data Fig. 8: Combination of clinical drugs elicits FL-GSDME-mediated pyroptosis.
From: Full-length GSDME mediates pyroptosis independent from cleavage
In this figure, HeLa cells were treated with different combination of reagents as indicated for 1 h to show GSDME PARylation, 2 h to show GSDME conformational change, 3 h to detect GSDME PM targeting, 4 h to indicate GSDME oxidation, 5 h to show pyroptotic morphology. a, different reagents were used to treat cells (5-Fluorouracil, 50 μM; Actinomycin D, 2 μg/ml; Bleomycin, 50 μM; Busulfan, 200 μM; Cisplatin, 20 μg/ml; Cisprofloxacin, 1 mM; Doxorubicin, 20 μM; Hydroxyurea, 2 mM; Mitoxantrone, 12.5 μM; Oxaliplatin, 15 μM; Paclitaxel, 10 μg/ml; Temozolomide, 200 μM; Topotecan, 20 μM; and VP-16, 100 μM) for 9 h, the cell morphologies were observed. b, RSL3 (100 nM) combined with different reagents, including MMS (methyl methanesulfonate, 1 mM), MNNG (N-methyl-N’-nitro-N’-nitrosoguanidine, 0.3 mM) and carmustine (0.25 mM)), were used to treated cells, the pyroptotic morphologies were observed. RSL3 combined with lower dose of UVC irradiation (50 mJ cm-2) was used as a positive control. c, cells transfected with GSDME-Strep-Flag were treated with RSL3 combined with different reagents or UVC irradiation (50 mJ cm-2), GSDME PARylation was determined. d, cells transfected with GSDME-mut2-HA were treated with RSL3 combined with different reagents or UVC irradiation (50 mJ cm-2), the GSDME conformation change was determined. e-f, cells were treated with RSL3 combined with different reagents or UVC irradiation (50 mJ cm-2), the GSDME oxidation (e) and GSDME PM targeting were determined (f). g, Representative flow cytometry plots illustrated the distribution of natural killer (NK) cells, perforin (PFN) and interferon-gamma (IFN-γ) expression in CD8+ T cells, along with IFN-γ expression in NK cells within the tumour microenvironment. All western blots were repeated at least two times and one of them is shown.
