Figure 1.
Figure 1. Effects of TK inhibitors on KIT phosphorylation in neoplastic cells. (A-B) KIT phosphorylation in HMC-1.1 cells (A) and HMC-1.2 cells (exhibiting KIT D816V) (B) after incubation in control medium, imatinib (1 μM), PKC412 (1 μM), or AMN107 (1 μM) for 4 hours. (C-D) KIT phosphorylation in Ton.Kit.wt cells (C) and Ton.Kit.D816V.27 cells (D) after incubation in control medium (control), imatinib (1 μM), PKC412 (1 μM), or AMN107 (1 μM) for 4 hours. Prior to drug exposure, Ton.Kit.wt cells and Ton.Kit.D816V.27 cells were kept in doxycycline for 24 hours to induce expression of KIT. In case of the Ton.Kit.wt clone, cells were also exposed to SCF (100 ng/mL, 4 hours) to induce KIT phosphorylation (p-KIT). In all cells, immunoprecipitation was conducted using the anti-KIT mAb SR-1. Western blotting was performed using the antiphospho mAb 4G10 for p-KIT detection and the anti-KIT mAb 1C1 for detection of total KIT protein.

Effects of TK inhibitors on KIT phosphorylation in neoplastic cells. (A-B) KIT phosphorylation in HMC-1.1 cells (A) and HMC-1.2 cells (exhibiting KIT D816V) (B) after incubation in control medium, imatinib (1 μM), PKC412 (1 μM), or AMN107 (1 μM) for 4 hours. (C-D) KIT phosphorylation in Ton.Kit.wt cells (C) and Ton.Kit.D816V.27 cells (D) after incubation in control medium (control), imatinib (1 μM), PKC412 (1 μM), or AMN107 (1 μM) for 4 hours. Prior to drug exposure, Ton.Kit.wt cells and Ton.Kit.D816V.27 cells were kept in doxycycline for 24 hours to induce expression of KIT. In case of the Ton.Kit.wt clone, cells were also exposed to SCF (100 ng/mL, 4 hours) to induce KIT phosphorylation (p-KIT). In all cells, immunoprecipitation was conducted using the anti-KIT mAb SR-1. Western blotting was performed using the antiphospho mAb 4G10 for p-KIT detection and the anti-KIT mAb 1C1 for detection of total KIT protein.

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