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. 2019 Apr 20;24:101206. doi: 10.1016/j.redox.2019.101206

Fig. 11.

Fig. 11

Effects of post-treatment with novel derivatives of vitamin D3 and lumisterol on Nrf2 targeted antioxidant proteins following UVB-irradiation. Keratinocytes were exposed to UVB intensities of 50 mJ/cm2 and immediately treated with the indicated hydroxy-derivatives of vitamin D3 or lumisterol (100 nM), or ethanol vehicle, for various times depending on the detection assay. Catalase (CAT) (A) and HO-1 (B) levels were determined at 3 h, while MnSOD (C) was determined 1 h post UVB irradiation. Cells were fixed and stained with anti-CAT, HO-1, or MnSOD antibody, imaged with a fluorescence microscope and analysed using the Cytation 5 reader and Graph Pad Prizm. Data are shown as the relative fluorescence intensities of cells treated with the hydroxy-derivatives compared to irradiated, ethanol-treated control cells. Keratinocytes stained with CAT, HO-1, or MnSOD antibody exhibited green fluorescence while nuclear staining with propidium iodine caused red fluorescence. Data are mean ± SD. The statistical significance of differences was evaluated by the student t-test, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 for all conditions.