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. 2011 May;178(5):2344–2356. doi: 10.1016/j.ajpath.2011.01.020

Figure 4.

Figure 4

KIT D816V induces expression of OSM in Ba/F3 cells. A: Ton.Kit.D816V cells kept with IL-3 were transfected with plasmids pGV-OSM-luc and pRL-TK and then kept in the absence [control (Co)] or presence (Doxy) of doxycycline for 24 hours. Luciferase activities were then measured. OSM promoter activity was normalized to Renilla luciferase activity and reported as % of control ( = cells kept in the absence of doxycycline). B: Ton.Kit.wt [wild type (WT)] and Ton.Kit.D816V cells (D816V) kept in the absence of IL-3 were cultured without [control (Co)] or with doxycycline for 16 hours. In addition, cells were incubated with or without stem cell factor (SCF). Expression of OSM was then determined by quantitative real-time PCR. C: Ton.Kit.D816V cells kept in the absence of IL-3 were cultured in the absence [control (Co)] or presence of doxycycline. Expression of OSM was then determined by immunoblotting. Actin served as a loading control. D: Ton.Kit.D816V cells kept with IL-3 were cultured in the absence (Control) or presence (Doxy) of doxycycline and were treated with various concentrations of midostaurin as indicated for 16 hours. Expression of OSM was then determined by quantitative real-time PCR. All results represent the mean ± SD of three independent experiments. *P < 0.05.