Fig. 1.
Ligand-independent phosphorylation of FLT3/ITD in 32D cells.
Total cellular protein extracts derived from 1 × 107parental 32D cells and 32D subclones transduced with the pBabePuro vector, pBabePuro-FLT3, and pBabePuro-FLT3/ITD were immunoprecipitated with anti-FLT3 antibody. Cells were incubated without (lanes 1, 3, 5, 7) and with (lanes 2, 4, 6, 8) FL (100 ng/mL) for 5 minutes before extracts were made. The immunoprecipitates were resolved by 8% SDS-PAGE and subjected to immunoblot analysis with antiphosphotyrosine antibody 4G10 (top panel). The same membrane was stripped and reprobed with anti-FLT3 antibody (bottom panel).
