Figure 10

Impact of PKA inhibition on Rap1 activation and bradykinin-induced IL-8 release. Cells were treated for 30 min without (Basal) or with 100 μM Rp-8-CPT-cAMPS. In A, cells were first incubated with 100 μM 8-pCPT-2'-O-Me-cAMP or 500 μM 6-Bnz-cAMP for 5 min followed by measurement of GTP-loading of Rap1 as described in Material and Methods. Shown is a representative immunoblot. Alternatively, cells were stimulated with 10 μM bradykinin alone or in combination with 100 μM 8-pCPT-2'-O-Me-cAMP or 500 μM 6-Bnz-cAMP for 18 hrs (B). IL-8 release was then assessed by ELISA. Results are expressed as mean ± SEM of separate experiments (n = 3-7). *P < 0.05, **P < 0.01, compared to unstimulated control, ^§ P < 0.05 compared to basal condition.