Fig. 1

Radiotherapy induces GSDME-dependent pyroptosis in NPC cells through the intrinsic mitochondrial apoptotic pathway. A-B Six NPC cell lines were treated with or without 6 Gy irradiation. 72 h later, (A) cell morphological changes and (B) LDH release were measured. C Typical processes of cell pyroptosis, apoptosis and survival in HNE1 cells are shown. Brightfield and fluorescent images of stable EGFP-overexpressing NPC cells were taken at specific time-points post-6 Gy irradiation. D Percentage of pyroptotic, apoptotic, and viable cells in six NPC cells lines within 72 h after 6 Gy irradiation is indicated. E GSDME silencing inhibited pyroptosis (left), LDH release (upper right), and cell death (bottom right) in HONE1 cells at 72 h after 6 Gy irradiation. F Western blotting revealed the levels of indicated proteins in six NPC cell lines at 72 h after 6 Gy irradiation. Cyto C: cytochrome c. G Cell morphological changes (top), LDH release assay (bottom left), live cell imaging (bottom right), and (H) the levels of indicated proteins were assessed in HONE1 cell lines at the indicated time points after irradiation (6 Gy). (I) Cell morphological changes (top), LDH release assay (bottom left), live cell imaging (bottom right), and (J) the levels of indicated proteins were assessed in HONE1 cell lines at 72 h after the indicated irradiation dose. GAPDH was used to normalize the amount of protein loaded. All data are presented as the mean ± SD of three independent experiments. ns, No significant difference. ***P < 0.001, ****P < 0.0001