American Society of Hematology

Figure 2

$Figure 2. Kinase-dependent localization of oncogenic Kit and PI3-K signaling molecules in lipid rafts. (A) Effect of imatinib treatment on the subcellular localization of oncogenic Kit in HMC-1 cells. Subcellular fractionation was performed as for Figure 1, using the HMC-1 cell line expressing the oncogenic Kit560V>G. Cells were incubated in imatinib containing media (1 μM) for 3 or 12 hours. Fractions were immunoblotted with antibodies against Kit and PY. Kit and phosphorylated Kit proteins are indicated by and , r, respectively. For quality control of the fractions, lipid raft and membrane fractions were verified by IB for Flotillin-1 and Na/K ATPase. (B) Effect of imatinib treatment on the localization of p85 in lipid rafts of HMC-1 cells. Fractions were immunoblotted with antibodies against p85, p110, PTEN, Akt, and PDK1.$

Kinase-dependent localization of oncogenic Kit and PI3-K signaling molecules in lipid rafts. (A) Effect of imatinib treatment on the subcellular localization of oncogenic Kit in HMC-1 cells. Subcellular fractionation was performed as for Figure 1, using the HMC-1 cell line expressing the oncogenic Kit^560V>G. Cells were incubated in imatinib containing media (1 μM) for 3 or 12 hours. Fractions were immunoblotted with antibodies against Kit and PY. Kit and phosphorylated Kit proteins are indicated by and , r, respectively. For quality control of the fractions, lipid raft and membrane fractions were verified by IB for Flotillin-1 and Na/K ATPase. (B) Effect of imatinib treatment on the localization of p85 in lipid rafts of HMC-1 cells. Fractions were immunoblotted with antibodies against p85, p110, PTEN, Akt, and PDK1.

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