Extended Data Fig. 9: Inhibition of GSDME-NT autophagic degradation improves chemotherapeutic response in vivo.
(a) Immunoblots of CT26 and B16 cells overexpressing mGSDME. (b) B16 mGSDME ATG4B(C74A)Tet-On cells were implanted into female C57BL/6 mice, with Dox and/or cisplatin (n = 6). (c) Immunoblots and analysis of mouse GSDME in tumours. (d) Immunoblots showing the subcellular localization of mouse GSDME-NT in tumours. (e) Representative immunofluorescence images and the percentage of negative nuclear HMGB1 cells in tumour sections treated as in (b). Scale bar: 100 µm. (f) Gating strategy of CD45 + , F4/80-, CD11B-, CD11C + DC cells. (g-h) Flow cytometry analysis of CD80 and MHC II expression in DCs (g), CD8 + T cells and IFN-γ, TNF production (h) in tumours treated as in (b). (i) Knockout efficiency of mouse GSDME. (j-k) Gsdme knockout CT26 mGSDME Flag–ATG4B(C74A)Tet-On cells were implanted in BALB/c mice and treated with Dox and/or cisplatin (n = 6). Tumour growth curves (j). Flow cytometric analysis of CD8 + T cells and IFN-γ, TNF-α production (k). (l) Growth of CT26 mGSDME Flag–ATG4B(C74A)Tet-On cells in BALB/c mice treated with anti-CD8 antibody. Mice received Dox and/or cisplatin (n = 6). (m) Knockdown efficiency of mouse TOLLIP. (n-q) B16 mGSDMETet-Tollip shRNA cells were implanted into C57BL/6 mice, with Dox and/or cisplatin (n = 6). Tumour growth curves (n). Immunoblots and analysis of mouse GSDME in tumours (o). Flow cytometry analysis of CD80 and MHC II of DCs (p), CD8 + T cells and IFN-γ, TNF production (q). Data are shown as Mean ± SD. n = 6 biologically independent mice. Two-way ANOVA with Tukey’s multiple-comparison test (b, j, l, n), two-tailed unpaired Student’s t test (c, d, o) or one-way ANOVA (e, g, h, k, p, q) were used for analysis.
