CellGeo analysis of genetically manipulated D16C3 cells. (A–A″) Drosophila D16C3 cells stained for Dia (A) and phalloidin (A′). (A″, inset) Dia at filopodia tips (left arrow) and cell cortex (right arrow). (B–B″) D16C3 cells stained for Ena (B) and phalloidin (B′). (B″, inset) Ena at filopodia tips (right arrow) and cell cortex (left arrow). (C–H) D16C3 cells expressing fluorescent actin and overlaid with the cell boundary (blue) and filopodia (red). Protrusions not meeting L_cr are in yellow. (C′–H′) Stills. Videos of broad protrusion dynamics (with filopodia removed) for cells in C–H in which every three frames are compared (lag 3). Cell bodies at time t (red) or t + 3 (blue) with areas of protrusion in white and retraction in black. (C and C′) Wild-type cell expressing actin alone. (D and D′) DiaΔDAD expression. (E and E′) Ena overexpression. (F and F′) Control cell treated with SK RNAi (against BlueScript plasmid). (G and G′) Dia knockdown. (H and H′) Ena knockdown. KD, knockdown; OE, overexpression; WT, wild type. Bars: (C, D, E, H) 5 µm; (A, A″ [inset], and F) 10 µm; (G) 15 µm.