Figure 3: The calcium signalling cascade mediates the ISO-increased rAAV transduction. The fold activation of rAAV transduction is indicated following preincubation with: 100µM PKI, 1µM calphostin C, 1µM EGTA, 50µM PD98059 (PD), 10µM SB203580 (SB), 0.1µM wortmannin or 20µM genistein with (A) or without 10-5 M ISO (B) in H9C2 cells cells compared with control. Data are meanħSEM, *p<0.05 (n=4). (C) The fold activation of rAAV transduction is indicated following preincubation with the indicated concentration of EGTA, 10µM BAPTA-AM, 0.1µM W7, 500 ng/ml cyclosporin A (CyA), 500µM calcineurin autoinhibitory peptides (CNIP), 1µM ionomycin in H9C2 cells compared with ISO alone or control. Data are meanħSEM, *p<0.01 (n=4) vs. ISO alone, **p<0.01 (n=4) vs. PBS control. (D) Inhibition of calcineurin by CNIP suppresses rAAV transgene expression as shown by RT-PCR analysis of lacZ gene expression. β-actin control expression in the cells is shown. (E) Abrogation of ISO-increased transduction of pseudotype rAAV1/2 vectors following CNIP incubation in H9C2 cells. Data are meanħSEM, *p<0.05 (n=4) vs. PBS control, **p<0.05 (n=4) vs. ISO alone.