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Copyright by Eunyoung Park 2007 - The University of Texas at Austin

Copyright by Eunyoung Park 2007 - The University of Texas at Austin

FIGURE 4.3. RETINOL

FIGURE 4.3. RETINOL DECREASES PI3K ACTIVITY BY INHIBITING PI3K AND PI INTERACTION. 93

FIGURE 4.3. RETINOL DECREASES PI3K ACTIVITY BY INHIBITING PI3K AND PI INTERACTION. Total protein was isolated from HCT-116 (A and C) and SW620 (B) cells treated with 10% FBS for 30 min after 24 h of serum starvation. Total PI3K was immunoprecipitated using p110 antibody as described in the Materials and Methods section. Immunoprecipitated PI3K was incubated with increasing concentrations of retinol and 10 μg PI for 10 min and then incubated with [γ- 32 P] ATP for 20 min (A and B). Immunoprecipitated PI3K was incubated with increasing concentrations of PI and 10 μg retinol for 10 min and then incubated with [γ- 32 P] ATP for 20 min (C). Total immunoprecipitated PI3K was detected by p110 antibody and used as an internal loading control. Following incubation the lipids were extracted and separated by TLC as described in the Materials and Methods Section. Data are reported as mean ± SEM, of three independent experiments. *Significantly different from control (P < 0.05). 94

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