Figure 8. Inhibition of PI 3-kinase does not affect CCL2 uptake and monocyte migration.

(A) Selected frames from time-lapse video microscopy (Movie S9) of migrating human monocytes recorded at 37°C. Monocytes were pretreated with 100 nM wortmannin for 30 min. The micropipette was dispending 100 nM CCL2-mCherry with constant backpressure. Gray (left column) corresponding phase image. Left panels phase images, middle panels CCL2-mCherry (ex 594 nm/em 620–680 nm), and right overlay. Time stamps are indicated in mm∶ss. (B and C) Analysis of directionality and velocity of monocytes: effect of PI 3-kinase inhibition. Tracks of monocytes (recorded as in Fig. 1 or Fig. 8A) either untreated (B) or pretreated with 100 nM wortmannin (C) were analyzed and plotted aligning their average directional vector with the y-axis (distance in µm). (D) Mean velocity of tracks shown in (B) and (C) were calculated using Metamorph software and median speed (MS) values are reported. Data from two independent observations, n = 16 (A) and n = 15 (B). MI = migratory index is the distance between the starting point and the end point divided by the total migrated distance.