Role of PKC, calcineurin, PI3-kinase and P2Y12 receptor in secretion and cofilin dephosphorylation in thrombin-activated platelets
| Inhibitors (conc.) . | Targeted proteins . | ATP secretion, % of control ± SD . | Inhibition of cofilin dephosphorylation . |
|---|---|---|---|
| DMSO | None | 100 ± 0 | No |
| Wortmannin (100 nM) | PI3-kinases | 100 ± 19 | No |
| Gö6976 (1 μM) | PKCα, βIand μ | 93 ± 8 | No |
| Gö6983 (2 μM) | PKCα, β, γ, δ and ξ | 2 ± 2 | No |
| CAID-11R (20 μM) | Calcineurin | 10 ± 6 | Yes |
| AR-C69931MX (2 μM) | P2Y12 | 91 ± 10 | No |
| Inhibitors (conc.) . | Targeted proteins . | ATP secretion, % of control ± SD . | Inhibition of cofilin dephosphorylation . |
|---|---|---|---|
| DMSO | None | 100 ± 0 | No |
| Wortmannin (100 nM) | PI3-kinases | 100 ± 19 | No |
| Gö6976 (1 μM) | PKCα, βIand μ | 93 ± 8 | No |
| Gö6983 (2 μM) | PKCα, β, γ, δ and ξ | 2 ± 2 | No |
| CAID-11R (20 μM) | Calcineurin | 10 ± 6 | Yes |
| AR-C69931MX (2 μM) | P2Y12 | 91 ± 10 | No |
Platelets were untreated or pretreated with inhibitors at 37°C for 20 minutes and then stimulated with thrombin (0.5 U/ml). ATP-secretion was measured using the Lumi-aggregometer, and cofilin dephosphorylation 30 seconds after thrombin stimulation was analyzed by immunoblotting as described in “Methods.”
conc. indicates concentration.