Figure 1.

CS recovers the decrease of filaggrin induced by TNF-α and IFN-γ treatment by inhibiting the activation of JNK. (A) HaCaT cells were incubated in the absence (medium alone) or presence of CS extract at the indicated concentrations for 48 h. Survival rate was measured by performing MTT‑based viability assay. Data are presented as a mean ± SD of three independent experiments and expressed as a relative ratio to the absorbance of untreated cells, which was set at 100%d. (B) HaCaT cells were preincubated in the absence and presence of CS at the indicated concentrations for 1 h. The cells were then incubated with 10 ng/mL TNF-α and IFN-γ for 48 h. The harvested cells were lysed, and filaggrin, loricrin and involucrin were analyzed by western blotting. (C) HaCaT cells were treated with TNF-α and IFN-γ in a time-dependent manner. The harvested cells were lysed and phospho-JNK was analyzed by western blotting. (D) HaCaT cells were pretreated for 1 h with or without 20 µM SP600125 (SP), after which the cells were treated with 10 ng/mL TNF‑α and IFN-γ for 48 h. The harvested cells were lysed and filaggrin was analyzed by western blotting. (E) HaCaT cells were pretreated for 1 h with or without CS extract, after which the cells were treated with 10 ng/mL TNF-α and IFN-γ for 1 h. The harvested cells were lysed and phospho-JNK was analyzed by western blotting. The membrane was stripped and reprobed with anti‑ERK2 or anti-JNK antibody as an internal control. Densitometric data are expressed as a mean ± SD and are presented relative to the negative control, which was set at 1 (right panels of B, C, D and E). *P < 0.05 and **P < 0.01 indicate a significant difference between the untreated and TNF-α and IFN-γ-treated group or between the TNF-α and IFN-γ-treated group and the CS or inhibitor-treated group.