Fig. 3: Blocking GSDMD-mediated pyroptosis alone did not decrease UVB-induced keratinocyte death.

A Photos of keratinocyte-specific Gsdmd cKO mice (Krt14^Cre/+-Gsdmd^flox/flox) and control mice (Krt14^+/+-Gsdmd^flox/flox) at 48 h after exposure or non-exposure to 430 mJ/cm² UVB irradiation (n = 4). B UVB skin damage of Krt14^Cre/+-Gsdmd^flox/flox mice and Krt14^+/+-Gsdmd^flox/flox mice was evaluated by scores (n = 4). C Representative images of H&E, TUNEL, MPO, MMP9, F4/80 staining in Krt14^Cre/+-Gsdmd^flox/flox mice and Krt14^+/+-Gsdmd^flox/flox mice after exposure or non-exposure to UVB irradiation. The statistic of epidermal thickness and average optical densities of TUNEL, MPO, MMP9, F4/80 positive staining were analyzed (n = 3). D Proteins of N-GSDMD, cleaved PARP, cleaved caspase-3, and p-MLKL in the epidermis of Krt14^Cre/+-Gsdmd^flox/flox mice and Krt14^+/+-Gsdmd^flox/flox mice were detected by western blotting assay. The relative protein levels of the interested proteins were compared with ACTB as quantification (n = 3). E Flowchart of DSF treatment or vehicle treatment for acute UVB skin injury model in WT mice (48 h after 430 mJ/cm² UVB irradiation). F Representative photos of mice receiving DSF or vehicle treatment at 48 h after UVB exposure. G UVB skin damage of DSF or vehicle treated mice was evaluated by scores (n = 8). H Representative images of H&E, TUNEL, MPO, MMP9, and F4/80 staining in DSF or vehicle treated mice after UVB irradiation. Statistical analysis of epidermal thickness and average optical densities of TUNEL, MPO, MMP9, F4/80 positive staining were shown (n = 8). I Proteins of GSDMD-FL, N-GSDMD, cleaved PARP, cleaved caspase-3, and p-MLKL in the epidermis of DSF or vehicle treated mice were detected by western blotting assay. The relative protein levels of the interested proteins were compared with ACTB as quantification (n = 3). The scale bar in black is 100 μm for 10× magnification in C and H. *P < 0.05. **P < 0.01. ***P < 0.001. ns not significant. DSF disulfiram. GSDMD-FL GSDMD full length. N-GSDMD N-terminal GSDMD. p-MLKL phosphorylated MLKL.