PD98059 inhibits GM-CSF mRNA stabilization in eosinophils activated by TNF-α plus Fn.
Eosinophils were incubated for 30 minutes with medium alone, 50 μM PD98059 (PD), or 10 μM SB203580 (SB). Then cells were activated for 5 hours with TNF-α + Fn (T + F) or medium alone (Resting) prior to transfection with GM-CSF mRNA by particle-mediated gene transfer. (A) At the indicated time points, equal numbers of cells were harvested, and total RNA was quantitatively isolated and Northern blotted with 32P-labeled GM-CSF or β-actin cDNA probes. Signals were visualized by using a PhosphorImager. (B) Radioactive signals were quantified and normalized to β-actin mRNA and plotted versus time. Each point is the mean (SD) of 3 experiments with 3 different donors.
