FLT3/ITD blocks 32D cell morphologic and functional differentiation to granulocytes.
32D/pBabe, 32D/FLT3, and 32D/FLT3/ITD cells were washed and transferred from medium containing IL-3 (1 ng/mL) to medium containing G-CSF (20 ng/mL) for 9 days. Every other day, fresh G-CSF–containing medium was added to cells to replace the old medium. (A) Cytospins were prepared on days 0 and 9. The cytospins were subjected to Wright-Giemsa staining followed by light microscopy. Original magnification, × 100. (B) Total cellular RNA from 32D/pBabe (lanes 1-6), 32D/FLT3 (lanes 7-12), and 32D/FLT3/ITD (lanes 13-18) cells was prepared from 1 × 107cells after 0, 1, 3, 5, 7, and 9 days in G-CSF. RNA (15 μg) from each time point was then subjected to Northern blotting with MPO, lysozyme, and actin cDNA probes.
