Figure 5

CypA expression in target cells. (A) CypA mRNA levels, expressed as copies per 1000 copies of GAPDH mRNA, were determined by real-time PCR using primers and probes shown in Table 2. Results are the mean ± SEM for 3 independent experiments using different cell pellets. (B) CypA protein expression. Equal amounts of soluble cell protein were electrophoresed into 10% SDS-PAGE gels, and transferred to PVDF membranes. The blots were probed sequentially with a rabbit anti-cypA antibody and a IRDye 800CW-conjugated donkey anti-rabbit IgG antibody. The blots were scanned using an Odyssey Infrared imaging system, and fluorescence intensity was analysed using Odyssey application software. Lysates from P4 cells were included in each gel, and the intensities of the CypA bands in the other samples on the same gel were expressed relative to that observed for P4 cells. The results presented are the mean ± SEM for 4 independent experiments, all performed using different cell suspensions.