Secreted factors from Arp2/3-depleted cells cause nonautonomous effects on chemotaxis. (A) Wind rose plots showing NS and 2×KD cells in NS CM or 2×KD CM in EGF chemotaxis. (B) Western blots showing phospho-AKT (p-AKT), total AKT, and GAPDH levels in NS and 2×KD cells upon EGF stimulation at the indicated time points. Representative blot from three experiments is shown. Fold changes in band intensity averaged across the three experiments (normalized to loading control and experimental control) are shown below each band. (C) Wind rose plots showing NS and 2×KD cells in NS CM or 2×KD CM during PDGF chemotaxis. (D) Blot showing p-AKT, total AKT, and GAPDH levels in NS and 2×KD cells upon EGF stimulation at the indicated time points. (E) Wind rose plots showing 2×KD cells in EGF chemotaxis, before (right) and after (left) switching from 2×KD CM to NS CM. 2×KD cells were plated in a chemotaxis chamber and serum starved overnight before carrying out the EGF chemotaxis experiment in the presence of 2×KD CM. After 4 h, 2×KD CM was switched to NS CM, and EGF chemotaxis was continued for another 4 h. Compass parameter and forward migration index are indicated on the graphs in A, C, and E; numbers in parentheses show 95% CI.