Involvement of PI3K pathway in SCF-induced cell migration.

(A) Association of PI3K p85 subunit with KIT was lost in Y719F. Total cell lysates prepared as described in Figure 4 were immunoprecipitated with anti-KIT antibody and blotted with anti-p85 antibody. The membrane was reblotted with anti-KIT antibody. (B) Effects of PI3K inhibitors on SCF-induced cell migration. Two different inhibitors, wortmannin or LY294002, were preincubtaed with BAF3/KIT cells and induced for migration with 30 ng/mL SCF. Migration without inhibitors was calculated as 100%. The results are shown as the means ± SD of triplicate cultures. (C) Dominant-negative PI3K inhibits SCF-induced Ca²⁺ influx. BAF3/KIT cells were stably transfected with dominant-negative PI3K (Δp85) and analyzed for SCF-induced Ca²⁺ influx (SCF; 100 ng/mL). (D) Dominant-negative PI3K inhibits SCF-induced cell migration. BAF3/KIT cells stably transfected with Δp85 or mock vector were analyzed for SCF-induced cell migration (SCF; 30 ng/mL). The asterisk represents a significant difference from a control (Mock) value (P < .01). Similar results were obtained from 3 independent experiments.