Fig. 6: ZBP1 is a key molecule in UVB-induced keratinocyte PANoptosis.
A Constitutive proteins related to PANoptosome including ZBP1, AIM2, NLRP3, NLRP12, RIPK1, RIPK3, p-RIPK3, caspase-8, cleaved caspase-1, FADD, and ASC in mice epidermal lysates were detected by western blotting assay at 48 h after 430 mJ/cm2 UVB irradiation. B Proteins of N-GSDMD, cleaved PARP, cleaved caspase-3, p-MLKL, and ZBP1 in human primary keratinocytes were detected by western blotting assay after UVB irradiation. C Representative images of immunohistochemical staining of ZBP1 in human skin samples from sun-exposed and non-exposed areas. Statistical analysis of average optical density of ZBP1 positive staining was shown (n = 12). D Representative photos of WT mice and Zbp1−/− mice at 48 h after 430 mJ/cm2 UVB exposure. E UVB skin damage of WT mice and Zbp1−/− mice was evaluated by scores (n = 5). F Representative images of H&E staining and TUNEL staining of skin sections from WT mice and Zbp1−/− mice after UVB irradiation. The statistic of epidermal thickness and average optical density of TUNEL positive staining were analyzed (n = 5). G Representative images of MPO, MMP9, and F4/80 immunohistochemical staining in WT mice and Zbp1−/− mice after UVB irradiation. The statistic of average optical densities of MPO, MMP9, and F4/80 positive staining were analyzed (n = 5). H Proteins of ZBP1, N-GSDMD, cleaved PARP, cleaved caspase-3, and p-MLKL in the epidermis of WT mice and Zbp1−/− mice were detected by western blotting assay. The relative protein levels of the above proteins were compared with ACTB as quantification (n = 3). The scale bar in black is 100 μm for 10× magnification in C, F, and G. The scale bar in yellow is 20 μm for 40× magnification in C and F. *P < 0.05. **P < 0.01. ns not significant. N-GSDMD N-terminal GSDMD. p-MLKL phosphorylated MLKL.
