Figure 6
Figure 6. S14161 inhibited PI3K activity. (A) PI3K activity analysis in an in vitro cell-free system. Increasing concentrations of S14161 were incubated with the PI3K isoforms α, β, γ, and δ, respectively. Activity of each kinase was determined with HotSpot technology as described in “Kinase activity in cell-free assay.” The results of 3 independent experiments are presented. Exp1, Exp2, and Exp3 indicate experiments 1, 2, and 3, respectively. (B) PI3K activity analysis after S14161 treatment in vivo. OPM2 cells were treated with S14161. Cell lysates were prepared and used for PI3K immunoprecipitation. PI3K activity was analyzed by ELISA as described in “Immunoprecipitation of P13K and P13K activity evaluation by ELISA.” Abs 450, absorbance at 450 nm of wavelength; S1, S14161; and LY, LY294002.

S14161 inhibited PI3K activity. (A) PI3K activity analysis in an in vitro cell-free system. Increasing concentrations of S14161 were incubated with the PI3K isoforms α, β, γ, and δ, respectively. Activity of each kinase was determined with HotSpot technology as described in “Kinase activity in cell-free assay.” The results of 3 independent experiments are presented. Exp1, Exp2, and Exp3 indicate experiments 1, 2, and 3, respectively. (B) PI3K activity analysis after S14161 treatment in vivo. OPM2 cells were treated with S14161. Cell lysates were prepared and used for PI3K immunoprecipitation. PI3K activity was analyzed by ELISA as described in “Immunoprecipitation of P13K and P13K activity evaluation by ELISA.” Abs 450, absorbance at 450 nm of wavelength; S1, S14161; and LY, LY294002.

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