Figure 3.

(A) Cytospin preparations of HEL cells and SET-2 cells were stained overnight with an anti-pSTAT5 antibody using immunocytochemistry. Scale bar: 20 µM. (B) HEL cells and SET-2 cells were stained with an anti-pSTAT5 Alexa-647 antibody for 30 min at room temperature and intracellular expression levels were analyzed by flow cytometry. (C) Nuclear (NE) and cytoplasmic (CE) fractions of HEL or SET-2 cells were analyzed for expression of phosphorylated STAT5 (pSTAT5), total STAT5a and total STAT5b by Western blotting. Antibodies against RAF-1 (cytoplasm) and TOPO-1 (nucleus) were used as fraction controls. The columns show the densitometry for pSTAT5. Uncropped blots are shown in Figure S1.