Fig. 8.

mRNA expression of MKP-1, GRα and translocation of GRα. Neutrophils were pretreated with salmeterol and FP or in combination for 15 min and then activated for 30 min for mRNA level of MKP-1 and GRα. Total RNA was isolated and subjected to RT-PCR using specific primers for MKP-1 (a) or for GRα (b). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) served as loading housekeeping control gene. Lower graphs a and b depicted for quantitative expression of MKP-1 and GRα mRNA, as a ratio to GAPDH mRNA. Representative results of three independent experiments are shown. c Neutrophils were pretreated with salmeterol or FP or in combination for 90 min and then activated with CSM (0.06 OD) for 30 min to determine GRα cytoplasmic and nuclear expression at protein levels. β-Actin and Lamin A served as loading controls from cytoplasm and nuclear fractions, respectively. Representative results of three independent experiments are shown. S and FP indicated for salmeterol and FP, respectively