Figure 1.
The abundance and kinase activity of PKCλ and PKCζ in mice with liver-specific PKCλ deficiency. (a) Total homogenates prepared from the liver of 20-week-old PKCλ+/+ mice were subjected to three sequential immunoprecipitations with antibodies to PKCλ (depleted PKCλ) or to PKCζ (deplete PKCζ). The resultant supernatants were subjected to immunoprecipitation with antibodies to PKCλ, to PKCζ, or with antibodies that recognize both PKCλ and PKCζ (Pan), and the precipitates were subjected to immunoblot analysis with antibodies that recognize both PKCλ and PKCζ (Pan). (b) Total homogenates prepared from the liver of 18-week-old PKCλ+/+ (WT); PKCλ+/+, Alb-Cre (WT-Cre); PKCλlox/lox (λlox/lox); or PKCλlox/lox, Alb-Cre (L-λKO) mice were subjected to immunoprecipitation with antibodies to PKCλ or to PKCζ, and the resulting precipitates were subjected to immunoblot analysis with corresponding antibodies. (c) Total homogenates prepared from the liver of λlox/lox or L-λKO mice were subjected to immunoprecipitation (IP) with antibodies to PKCλ or to PKCζ, and the resulting precipitates were assayed for kinase activity. (d) Extracts of the indicated tissues prepared from λlox/lox or L-λKO mice were subjected to immunoprecipitation with antibodies to PKCλ or to PKCζ, and the resulting precipitates were subjected to immunoblot analysis (IB) with corresponding antibodies. Data in a, b, and d are representative of at least three mice of each genotype; those in c are mean ± SEM of values from six mice. *P < 0.01 for the indicated comparison (ANOVA).