Fig. 2. Proliferation and high activation of signaling pathways are restored in
previously underexpressing clones that were retransduced to increase
expression of Mpl. (A) Proliferation assay. Cells were incubated with the
indicated concentration of TPO for 48 hours and cell proliferation was
quantitated by [3H]thymidine incorporation. The results shown are
the means±s.d. of triplicate experiments. (B) Western blots analysis of
signaling pathways activated in clones stimulated by TPO. Cells were washed
three times, deprived of cytokines for 3 hours, and then stimulated with 50
ng/ml of TPO for 15 minutes. Activation of STAT-5, ERK and AKT were determined
as described in Fig. 1. ns, no
stimulation.
