Fig. 5

The effects of daphnetin on p65 phosphorylation and nuclear translocation in M5-stimulated HaCaT keratinocytes. HaCaT keratinocytes were treated with daphnetin (Dap, 20 μM) for 24 h, and then stimulated with M5 (5 ng/ml) for 30 min. a Cells were harvested and total protein was extracted for Western blotting assay to detect p65 and p-p65 protein levels. β-actin served as an internal reference. *P < 0.05. b Cells were fixed and IFA was carried out to detect p65 (red) distribution. Nuclei were stained with Hoechst dye 33,258 (blue). Bar, 50 μm. DMSO (Ctl), DMSO + M5 (M5), daphnetin + M5 (M5 + Dap)