Fig. 3

The effects of GSDMD deletion on CNS of 5×FAD mice at the peak stage. a-d MWM test for 5×FAD and Gsdmd^−/−5×FAD mice at 6 months of age. Data are presented as representative swimming traces (a), escape latency in the training phase (b), number of crossing platform (c) and time spend in the target quadrant during the probe test (d) (n = 7/group). e-f Y-maze test for 5×FAD and Gsdmd^−/−5×FAD mice at 6 months of age. Data are presented as the bouts of entering target arm (e) and the percentage of duration in target arm (f) (n = 7/group). g-h The discrimination index of NOR (g) and NLR (h) tests for 5×FAD and Gsdmd^−/−5×FAD mice at 6 months of age (n = 7/group). i-l Immunofluorescence staining and quantification of Aβ plaques (Thioflavin S) (i,k) and IBA1 (j,l) in the hippocampus and cortex of 5×FAD and Gsdmd^−/−5×FAD mice at 6 months of age (n = 5/group). Scale bar = 200 μm. m-o ELISA analysis of IL-1β (m), IL-6 (n), and TNF-α (o) in the brain tissue of 5×FAD and Gsdmd^−/−5×FAD mice at 6 months of age (n = 5/group). p-r Flow cytometry analysis of immune cells (CD45^high), including infiltrating T cells (CD45^highCD8⁺ and CD45^highCD4⁺), myeloid cells and activated microglia (CD45^highCD11b⁺), in the brain tissue of 5×FAD and Gsdmd^−/−5×FAD mice at 6 months of age (n = 5/group). Data are presented as a representative plot (p), quantified percentages (q), and absolute cell numbers (r). Data are pooled from three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Error bars show means ± SEM. One-way ANOVA for B. Unpaired t test for c-h, k-o. Two-way ANOVA with Sidak’s multiple comparisons test for q, r