Figure 2

NHERF1 deletion leads to increased stability of PDGF-stimulated p-Akt signal in mouse embryonic fibroblasts. (a) Genotyping and Na⁺/H⁺ exchanger regulatory factor 1 (NHERF1) expression of mouse embryonic fibroblast (MEF) cells with NHERF1^+/+ and NHERF1^-/- backgrounds. Left panel: duplex PCR was conducted to evaluate the genotypes, using the genomic DNA extracted from MEFs. Wild-type (WT) and knockout (Neo) alleles were expected to generate 1.4-kilobase and 2.4-kilobase products, respectively. Predicted genotypes are indicated. Right panel: NHERF1 expression in MEFs as measured by immunoblotting. Asterisks indicate bands from nonspecific reactivity to mouse NHERF1 antibody. (b) MEFs (NHERF1^+/+ or NHERF1^-/-) were serum starved and stimulated with platelet-derived grwoth factor-BB (0.5 ng/ml) for indicated periods before cells were harvested for phospho-Akt (p-Akt; Ser473) measurement. Total Akt was also measured to verify equivalent loading.