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Fig. 5 Pt accumulated nuclear Nrf2 levels leading to HO-1, ?-GCLC expressions in HaCaT cells. 5 ?M Pt was treated to the cells. (A) The immunofluorescence method was used to examine (magnification × 200) the nuclear localization of Nrf2 protein. (B?C) The Western blot method determined the expression of nuclear Nrf2 (0?4 h) (B) or HO-1 and ?-GCLC proteins (0?8 h) (C). (D?E) Cells were first treated with Pt (0?5 ?M for 24 or 0.5 h), followed by 3 J/cm2 UVA-irradiation or not for the indicated time. The Western blot method determined the expressions of Nrf2, Keap-1, p-Nrf2 proteins (D). The Nrf2/Keap-1 ratio data were denoted as the fold-difference over untreated control (E). ?-actin functioned as a loading control protein. Results were denoted as mean ± SD of three or more independent experiments. Statistical significance was considered as **p < 0.01, ***p < 0.001 compared to untreated control and #p < 0.05, ###p < 0.001 compared to UVA-irradiated cells.