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Fig. 7. DECA disrupts the interaction between PKC-ß and RACK-I. (A) Subconfluent melanocyte cultures were treated with DECA or vehicle alone for 30 minutes. Then the cultures were UV irradiated (UVB, 12 mJ/cm2) to activate DECA and treated with 10–7 M TPA for an additional 90 minutes. Then cells were harvested and equal amounts of proteins from DECA- or vehicle-treated cells were subjected to immuno-precipitation using an antibody against RACK-I, followed by immunoblot analysis using antibodies against PKC-ß and RACK-I, as described for Fig. 5. Densitometric analysis of each band was performed. When the ratio of PKC-ß to RACK-I was set arbitrarily at 1.0, the ratio in DECA-treated samples was 0.25. (B) Subconfluent melanocyte cultures were treated with DECA or vehicle alone, UV-irradiated to activate DECA and then treated with 10–7 M TPA as described above. Cells were harvested and tyrosinase activity was determined as previously described (Pomerantz, 1964). Statistical analysis using a paired Student's t-test was performed.
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