Fig. 2

PI3 kinase association is necessary for the ligand-independent activation of secondary mutation of KIT. a, b Ba/F3 cells stably expressing KIT mutants were washed and starved in RPMI 1640 medium for 4 h in the presence of PI3 kinase inhibitor Copanlisib (50 nM). After stimulation with 100 ng/ml SCF for 2 min and cell lysis, KitC1 was used to precipitate KIT from cell lysate. pY antibody 4G10 and KitC1 were used to detect KIT activation. pAkt, Akt and β-actin antibody were used to detect Akt activation in total cell lysates. Signal intensity was quantified and calculated to show the relative KIT activation