Figure 3.

Fluvastatin prevents translocation of FLT3/ITD to the cell surface leading to intracellular accumulation of the immature form. (A) BaF3 FLT3/ITD cells were treated with 1μM fluvastatin for 24 hours. Afterward, surface or total FLT3/ITD was measured by incubating with CD135-PE for 30 minutes and analyzed by flow cytometry. The untreated cells are shown in red, fluvastatin-treated cells in blue, and total FLT3 from cells treated with fluvastatin and then permeabilized is shown in green. Cell-surface staining with IgG is shown in black for both the permeabilized and nonpermeabilized cells. (B) Cells treated with or without fluvastatin were adhered to poly-L-lysine slides and stained for plasma membrane (WGA-A350, blue) or FLT3 (Cy3, red). Slides were visualized by immunofluorescence microscopy using a Nikon TE2000 Eclipse microscope and 60 × 1.4 oil-immersion lens. (C) Kasumi cells were treated with 1μM fluvastatin for 24 hours and assessed for surface c-Kit expression. Cells stained with IgG are shown in black, untreated cells in red, and fluvastatin-treated cells in blue. (D) Western blot results show fluvastatin treatment led to a reduction in the amount of mature c-Kit expression in Kasumi-1 cells. Results are representative of at least 3 independent experiments.