Fig. 5.

ITI-214 inhibits ADP induced migration of BV2 cells in a dose dependent manner. BV2 cells were added to upper chamber of a 5 μm pore Boyden chamber with 100 μM ADP or vehicle in the lower chamber and incubated at 37°C with 5% CO₂ for 4 h. Indicated doses of ITI-214 were included in the upper chamber at time 0. (A) Raw fluorescence counts (relative fluorescence units or RFU), representing the cell count in the lower chamber, for the control conditions. The Vehicle condition is the minimum migration; there is no ADP present. The maximum migration is the 100 μM ADP condition. The average of each control condition in the left graph was used to normalize the data from the ITI-214 treated samples. The curve was fit to a 4-parameter logarithmic equation with the following constraints: top > 80% and bottom < 20%. The resulting IC50 is 0.16 μM. n = 8 (B) Dose dependent inhibition of ADP chemotaxis by P2Y12 inhibitors. AR-C 66096 (open circles; documented IC50 of 6.9 nM) and AR-C 69931 (black circles, documented IC50 of 0.4 nM) inhibit ADP induced migration in a Boyden chamber system.