Fig. 3.
Effect of PI 3-kinase inhibition on MAP-kinase activation.
MAP-kinase activation observed in human erythroid progenitors and UT7 cells stimulated with Epo is independent of PI 3-kinase activation. Lysates were prepared from human erythroid progenitors stimulated 10 minutes with Epo at 10 U/mL, without or with LY294002 or rapamycin or both compounds, and analyzed by Western blot with the use of anti-pERK, anti-pFKHRL1, anti-p70S6K, and anti-ERK2 antibodies. Lysates obtained from UT7 cells stimulated at various times in the presence or absence of LY294002 at 50 μm were analyzed for MAPK activation with the use of an anti-pERK antibody. The blots were stripped and reprobed with an anti-ERK2 antibody.
