Fig. 3: GSDMD regulates the recruitment of neutrophils to the irradiated liver.

A, B Workflow and Proportion plot of immune cells. A Livers from Gsdmd^FL/FL -Ctrl, Gsdmd^FL/FL-ST and Gsdmd^∆Hep-ST group were used. 7 days after the last 30 Gy irradiation, the livers were digested and 10 × Genomics scRNA-seq was used to profile the cells; B UMAP plot of liver single CD45⁺ cells from three group mice, colored by the group. C Colors indicate the cell type annotated using Single R and manual annotation and validated by cell-type-specific gene expression. D Flow Cytometric analysis of liver-infiltrated neutrophils (FVD^-CD45⁺CD11b⁺Ly6G⁺) of Gsdmd^FL/FL and Gsdmd^∆Hep mice. Gsdmd^FL/FL vs Gsdmd^∆Hep: ST, p = 3.1E-09; LT, p = 0.002. E Neutrophil infiltrationwas analyzed by immunohistology staining (left, scale bars, 200×, 50 μm) with antibodies against MPO, and following morphometric quantification (right). D, E n = 3/group, Ctrl; n = 5/group, ST/LT. Gsdmd^FL/FL vs Gsdmd^∆Hep: ST, p = 1.218E-07; LT, p = 2.86E-07. F, G Detection of the neutrophil infiltration in liver of Gsdmd^∆Hep mice treated with or without AAV9-GSDMD-FL adeno-associated virus. F Representative plots (left) and statical analysis (right) of the percentage of CD11b⁺Ly6G⁺ cells with the flow cytometry. **p = 0.005. G MPO staining images (left) and quantification (right). ****P < 0.0001. F, G n = 4/group. H Quantification of neutrophil migration in response to Gsdmd^FL/FL and Gsdmd^∆Hep-hepatocytes pretreated with irradiation or not. Each sample was from 3 biologically independent replicates. ***P = 0.0003. I The depletion effect of neutrophils in liver were tested by flow cytometry in ST-RILD mice. ****P < 0.0001. J Representative liver HE images from ST-RILD mice (Scale bars, 200×, 50 μm). K Serum levels of ALT, AST, and ALB.****P < 0.0001. n = 5 mice/group for I–K. p values were using two-way ANOVA with Fisher’s LSD multiple comparisions test (D, E, H) and unpaired two-sided Student’s t test (F, G and I). No data were excluded from the analyses. Data are expressed as mean ± SEM. Source data are provided as a Source Data file.