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. 2002 Feb;22(3):965–977. doi: 10.1128/MCB.22.3.965-977.2002

FIG. 6.

FIG. 6.

Effect of disruption of Pik3r1 on downstream kinases from PI 3-kinase. (a) IGF-1-induced Akt activity in cells of each genotype. Cells were starved for 24 h and then stimulated with 10 nM IGF-1 for 5 min. Cell lysates were subjected to immunoblotting with anti-phospho-Akt (αphospho-Akt; top panel) antibody or immunoprecipitation with αAkt antibody. The immunoprecipitates were subjected to an immune complex kinase assay. In the bottom panel, each bar represents the mean ± standard deviation of the relative Akt kinase activity calculated from the results of three independent experiments. *, P value of <0.01 for wild-type (Wild) versus heterozygous KO (Hetero) cells; **, P value of <0.05 for wild versus null cells. (b) IGF-1-induced p70S6K activity in cells of each genotype. After 20 min of stimulation with 10 nM IGF-1, cell lysates were subjected to immunoblotting with anti-phospho-p70S6K (αphospho-p70S6K; top panel) antibody or immunoprecipitation with αp70S6K antibody. The immunoprecipitates were subjected to an immune complex kinase assay. In the bottom panel, each bar represents the mean ± standard deviation of the relative p70S6K kinase activity calculated from the results of three independent experiments.