Clearance of UV photoproducts was assessed in whole skin biopsies as previously reported (Jarrett et al., 2014 (link)). Briefly, mice pre-treated (twice a day for 3 days) with vehicle or forskolin were depilated 4 days prior to radiation (7.5 kJ/m2). 1 cm2 skin biopsies were collected at the indicated time points after UV exposure, DNA was isolated by DNEasy kit (Qiagen, Germantown, MD) and quantified by Nanovue nanodrop (GE Biosciences, Pittsburgh, PA). 0.1 μg/well DNA was loaded onto nitrocellulose membranes using a slot blot apparatus. Membranes were processed as described (Wang, Wu, & Friedberg, 1991 (link)) and immunoblotted for [6,4]-photoproducts using a monoclonal antibody against this UV photolesion (Cosmo Bio, Tokyo, Japan). Band intensities were analyzed using ImageJ software.
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UV Photoproduct Clearance in Skin
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Corresponding Organization : Markey Cancer Center
Other organizations : Fujita Health University
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Variable analysis
independent variables
- Vehicle or forskolin treatment (twice a day for 3 days)
- UV radiation (7.5 kJ/m^2)
- Clearance of UV photoproducts in whole skin biopsies
- Time points after UV exposure
- DNA isolation and quantification method
- Slot blot apparatus and immunoblotting for [6,4]-photoproducts
- Positive control: Not explicitly mentioned.
- Negative control: Mice pre-treated with vehicle (instead of forskolin)
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