Fig. 6: MLKL-mediated HMGB1 secretion from keratinocyte facilitates DNCB-motivated CHS.
A HMGB1 in the DNCB-motivated ears of Mlkl cKO mice and control mice were detected by immunohistochemistry assay. B The protein level of HMGB1 in epidermis was detected by western blotting. Statistical analysis of the interested protein was shown (n = 3). C HMGB1 in the DNCB-motivated ears of wild type mice treated by intraperitoneal injection with TC13172 or solvent PBS were detected by immunohistochemistry assay. D The protein level of HMGB1 in epidermis was detected by western blotting. Statistical analysis of the interested protein was shown (n = 3). E HMGB1 in DNCB-motivated ears of wild type mice topically treated by necrostatin-1 or vehicle were detected by immunohistochemistry assay. F The protein level of HMGB1 in epidermis was detected by western blotting. Statistical analysis of the interested protein was shown (n = 3). G–I Mlkl cKO mice (Krt14Cre/+-Mlklflox/flox) and control mice (Krt14+/+-Mlklflox/flox) were intradermal injected with rHMGB1 (right ear) and PBS (left ear), respectively. The skin appearance (G), histology feature (H) and infiltration of MPO, MMP-9 and NE (I) were shown. J The protein levels of HMGB1 in nucleus, cell plasma and supernatant were detected by western blotting in the MLKL knockdown HaCaT cells in the presence or absence of TNF-α and IFN-γ. Statistical analysis of the interested protein was shown (n = 3). K HMGB1 secretion in supernatants of cultured HaCaT cells were determined by ELISA. Scale bar (A, C, E) represents 20 μm. Scale bar (H, I) represents 200 μm. ∗p < 0.05.
