Transduced Tat-dn-H-Ras blocks activation of endogenous Ras on IL-5 stimulation.

Human blood eosinophils were incubated with 100 nM Tat-dn-H-Ras for 30 minutes and then stimulated for 7 minutes with 1 nM IL-5. Resultant cell lysates were incubated with GST-Ras binding domain protein bound to glutathione agarose beads for 30 minutes at 4°C. After they were washed with lysis buffer, the mixture was boiled in sample buffer, separated by SDS-PAGE, and immunoblotted using anti-Ras antibody. Figure 3 is representative of 4 repetitions of this experiment.