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. 2024 Dec 24;13:RP101248. doi: 10.7554/eLife.101248

Figure 4. GSDMD depletion in neutrophils attenuates the development of skin inflammation in psoriasis.

(a) Representative images of western blot showing the expression of GSDMD in bone marrow-derived neutrophils of cKO and isotype control mice. (b) Macroscopic phenotypic representation of the dorsal skin in control and GSDMD-cKO mice treated with IMQ at day 4. (c) Disease severity of psoriasis induced by IMQ in mice as assessed by PASI score (n=7). (d) Representative images and statistical analysis of hematoxylin and eosin staining of the dorsal skin in control and GSDMD-cKO mice treated with IMQ at day 4 (n=5). Scale bar = 100 µm. (e) Quantitative PCR analysis of the relative mRNA expression of proinflammatory cytokines in the dorsal skin of control and GSDMD-cKO mice treated with IMQ at day 4 (n=4). Data were normalized to a reference gene, GAPDH. (f) ELISA analysis of IL-6 and IL-1β per 1 mg of the dorsal skin from control and GSDMD-cKO mice treated with IMQ at day 4 (n=5). (g) Representative images and statistical analysis of western blot analysis showing the expression level of GSDMD and its N-terminal fragments in dorsal skin of WT, cKO, and Gsdmd-/- mice treated with IMQ at day 4 (n=4). (h) Schematic representation of the use of anti-Ly6G antibody in the IMQ-induced psoriasis mouse model. (i) Macroscopic phenotypic representation and PASI score of cKO mice and control mice treated with IMQ and Ly6G antibody. cKO, conditional knockout; ELISA, enzyme-linked immunosorbent assay; IMQ, imiquimod; PASI, psoriasis area and severity index, WT, wild-type. Error bars show mean ± SEM. *p<0.05, **p<0.01, ***p<0.001. Data are representative of three independent experiments for (a, b, d, g, i).

Figure 4—source data 1. PDF file containing original western blots for Figure 4a and g, indicating the relevant bands and treatments.
Figure 4—source data 2. Original files for western blot analysis displayed in Figure 4a and g.

Figure 4.

Figure 4—figure supplement 1. Flow cytometry plots of neutrophil, and data analysis in single-cell RNA sequencing from the skin of IMQ-induced psoriasis-like mice.

Figure 4—figure supplement 1.

(a) Flow cytometry plots of neutrophil, isolated from dorsal skin of wild-type (WT) mice treated with imiquimod (IMQ). (b) Uniform Manifold Approximation and Projection (UMAP) of single cells isolated from the skin of IMQ-induced psoriasis-like mice (GSE165021), and feature plots showing expression of Il17a, Il6, Tnfa, Il1b, and Il18 (n=2). Data are representative of three independent experiments for (a).