Fig. 1: Methionine restriction promotes GSDME-mediated pyroptosis.
From: Methionine metabolite spermidine inhibits tumor pyroptosis by enhancing MYO6-mediated endocytosis
(A) Schematic depicting the workflow for model-building and the screening method. (B and C) Effects of individual amino acid deprivation on Dox-induced PI uptake (B) and LDH release (C) in GSDME-NTTet-On HONE1 and HCT116. EAA, essential amino acid; NEAA, non-essential amino acid. (D) Immunoblot of cleaved caspase-3 in Dox-treated GSDME-NTTet-On HONE1 and HCT116 cultured in normal or Met-omitted medium with or without ZDEVD-FMK (30 μM) treatment. (E and F) Effects of zDEVD-FMK on PI uptake (E) and LDH release (F) in Dox-treated GSDME-NTTet-On HONE1 and HCT116 cultured with normal or Met-omitted medium. All p-values comparing the uptake of PI and the release of LDH in cells subjected to Met deficiency, in response to Dox, with those in cells subjected to deficiency of other amino acids, are less than 0.0001. (G) Effects of MRD on serum Met level of BALB/c-nu/nu mice (n = 6). (H) Immunofluorescence of HMGB1 in GSDME-shTet-On HONE1 tumors of indicated groups. Scale bar: 100μm. (I) Quantification of nuclear HMGB1- cells shown in (H) (n = 6). Data are represented as mean ± SD. Two-way ANOVA with Bonferroni’s multiple comparisons test (B, C, E, F); Two-tailed Student’s t test (G); One-way ANOVA with Tukey’s multiple comparisons test (I). NS, not significant. The results are representative of three independent experiments (B–F). Source data are provided as a Source Data file.
