Fig. 5: Dexamethasone or necroptosis inhibitor necrostatin-1 inhibits necroptosis in TNF-α and IFN-γ- treated HaCaT cells. | Cell Death Discovery

Fig. 5: Dexamethasone or necroptosis inhibitor necrostatin-1 inhibits necroptosis in TNF-α and IFN-γ- treated HaCaT cells.

From: Necroptosis-mediated HMGB1 secretion of keratinocytes as a key step for inflammation development in contact hypersensitivity

Fig. 5

A, B The protein level and phosphorylation of RIPK1, RIPK3 and MLKL were detected by western blotting in the TNF-α and IFN-γ- treated HaCaT cells in the presence or absence of dexamethasone (A), as well as necrostatin-1 (B). Statistical analysis of the interested protein was shown (n = 3). Trypan blue staining (C) and PI staining (D) were used to determine the levels of cell death in TNF-α and IFN-γ- treated HaCaT cells in the presence or absence of dexamethasone or necrostatin-1. The photos of HaCaT cells in (C) are 20× magnification. And the threshold is 4–30 μm. Scale bar (D) represents 100 μm. ∗: p < 0.05. E, F Necroptosis was induced by the treatment of TNF-α (100 ng/ml), Smac mimetic compound (100 nM) and z-VAD-fmk (20 μM) (TSZ) in HaCaT cells for 12 h, and protein level and phosphorylation of RIPK1, RIPK3 and MLKL were detected by western blotting. Necrostatin-1 treatment was used to validate the inhibition of necroptosis. Statistical analysis of the interested protein was shown (n = 3). G PI staining was used to determine the levels of cell death in TSZ- treated HaCaT cells in the presence or absence of dexamethasone. Scale bar represents 100 μm.

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