Figure 3.

Responses to CCL2 but not fMLP are attenuated by DAG kinase and DAG lipase inhibitors in freshly isolated human monocytes. Effect of (a) DAG kinase inhibitor R59949 (30 μM) and (b) DAG lipase inhibitor RHC80267 (30 μM) on Ca²⁺ responses evoked by CCL2 (50 ng·ml⁻¹) in monocytes (n = 8). Averaged responses are shown in the presence of vehicle (closed circles) and inhibitor (open circles). (c) Bar chart showing effect of R59949 (30 μM) and RHC80267 (30 μM) on the peak Ca²⁺ response evoked by CCL2 (50 ng·ml⁻¹; n = 8). (d) Effect of R59949 (30 μM) and RHC80267 (30 μM) on freshly isolated monocyte transmigration to CCL2 (3 ng·ml⁻¹). *P < .05 versus vehicle and ^# P < .05 versus CCL2 alone (n = 8). Lack of effect of (e) R59949 (30 μM) or (f) RHC80267 (30 μM) on Ca²⁺ responses evoked by fMLP (10 μM) in monocytes (n = 6). Averaged responses are shown in the presence of vehicle (closed circles) and inhibitor (open circles). (g) Bar chart showing lack of effect of R59949 (30 μM) and RHC80267 (30 μM) on peak Ca²⁺ response evoked by fMLP (10 μM; n = 6). F ratio is the Ca²⁺ response as measured by the Fura‐2 emission intensity ratio when excited at 340 and 380 nm. Data in inhibition experiments are expressed as a percentage of the control response in the presence of vehicle alone