Fig. 8: GSDME inhibits STAT3 phosphorylation and S100a7a transcription by stabilizing OGT protein. | Nature Communications

Fig. 8: GSDME inhibits STAT3 phosphorylation and S100a7a transcription by stabilizing OGT protein.

From: Renal tubular GSDME protects cisplatin nephrotoxicity by impeding OGT-STAT3-S100A7A axis in male mice

Fig. 8

a,b Western blot analysis (a) and densitometric quantification (b) of GSDME-FL, S100A7A, p-STAT3, and STAT3 in primary WT PTCs treated with increasing concentrations of cisplatin (0, 5, 10, 20 μM) for 24 h (n = 4 biological replicates). c,d Western blot analysis (c) and densitometric quantification (d) of p-STAT3 and STAT3 in renal tissues from cisplatin-treated WT and Gsdme-KO mice (n = 12 biological replicates). e,f Western blot analysis (e) and densitometric quantification (f) of p-STAT3, STAT3, and S100A7A in primary WT and Gsdme-KO PTCs treated with or without cisplatin and/or Stattic (n = 4 biological replicates). g Co-IP assay examining the O-GlcNAcylation of STAT3 in cisplatin-treated primary WT and Gsdme-KO PTCs (n = 4 biological replicates). h Densitometric quantification of O-GlcNAcylation of STAT3, OGT, and OGA in panel (g) (n = 4 biological replicates). i RT-qPCR analysis of S100a7a mRNA expression in primary WT and Gsdme-KO PTCs treated with or without cisplatin and/or Thiamet G (n = 4 biological replicates). j Co-IP assay examining the O-GlcNAcylation of STAT3 in primary WT and Gsdme-KO PTCs treated with or without cisplatin and/or Thiamet G (n = 4 biological replicates). k Densitometric quantification of O-GlcNAcylation of STAT3, p-STAT3, and S100A7A protein levels in panel (i) (n = 4 biological replicates). l,m Western blot analysis (l) and densitometric quantification (m) of OGT in primary WT and Gsdme-KO PTCs treated with CHX for the indicated time (n = 4 biological replicates). n, o Co-IP assay examining the ubiquitination of OGT in primary WT and Gsdme-KO PTCs (n = 4 biological replicates). Data were expressed as means ± SEM. #P < 0.05, ##P < 0.01, ###P < 0.001 versus KO cisplatin group. *P < 0.05, **P < 0.01, ***P < 0.001 versus KO ctrl group. &P < 0.05, & &P < 0.01, & & &P < 0.001 versus WT ctrl group. P-values were determined by Student’s two-tailed t-test in (o) and one-way ANOVA (with LSD or Games-Howell post hoc tests) in (b, d, f, h, i, k). Source data are provided as a Source Data file.

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