Interactions of Lnk with FLT3-WT/ITD in hematopoetic cells. EOL-1 (A) or REH (B) cells were serum-starved for 16 hours (−) and treated with FL for 15 minutes (+). Cell lysates were subjected to pull-down by either anti-FLT3 Ab or normal rabbit Ig, and the precipitates were subjected to Western blotting analysis using Lnk Abs. Asterisk indicates the nonspecific bands and the heavy chain of Ig. Supernatant (SP) after immunoprecipitation from EOL-1, THP-1, and REH cells was included to show the position of endogenous Lnk. (C) 32D/FLT3-ITD cells were transfected with V5-tagged-Lnk. Lysates were precipitated with anti-V5 Ab, and the precipitates were subjected to Western blotting analysis using either FLT3 or V5 Abs.