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Figure 2


Fig. 2. CLIC4 is a downstream target of PKC{delta} in keratinocytes induced to differentiate by Ca2+. (A) Mouse keratinocytes were pre-treated with the indicated doses of PKC inhibitors for 1 hour, and then 0.5 mM Ca2+ was added in the medium for 24 hours prior to lysis and immunoblotting. (B) Mouse keratinocytes were infected with PKC{delta} DN Ad for 17 hours before the treatment with 0.5 mM Ca2+ for 24 hours. Actin was used as a loading control. Bis, bisindolylmaleimide I; Ro, rottlerin; Go, Go 6976. Similar data were obtained from two independent experiments. The numbers over the CLIC4 bands in A and B represent fold change determined by densitometry. (C) Purified recombinant CLIC1 or CLIC4 proteins were incubated with purified, kinase active recombinant PKC isoforms in appropriate reaction buffers with [{gamma}-32P]ATP, the reaction mixture was resolved by SDS-PAGE and then the phosphorylated proteins were visualized by phospho-imaging methods. Un, no recombinant PKC.





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