Figure 2.
Ena and Activated Dia Coexpression Drives Protrusion Dynamics Distinct from Either Alone
(A–O) D16 cells: arrows, filopodia; white arrowheads, lamellipodia; yellow arrowheads, Ena and Dia cortical colocalization. Transfection efficiency ranged from 10%–25% and expression levels were variable. Cells with midrange expression were used for all experiments.
(A–H′) D16 cells (Movie S1) expressing GFP-actin (A and A′), GFP-DiaΔDAD+RFP-actin (B–D′), mCh-Ena+GFP-actin (E–G′), or GFP-DiaΔDAD+mCh-Ena+GFP-actin (H and H′).
(I–K) Movie stills of GFP-DiaΔDAD (I and K) + mCh-Ena (J and K; Movie S3). Arrowhead, cortical colocalization in region without filopodia; white arrows, DiaΔDAD only filopodium.
(L and M) Mean filopodia number (L) and length (M) for Actin (n = 16), DiaΔDAD (n = 34), Ena (n = 31), or DiaΔDAD+Ena (n = 28). Error bars = 95% confidence interval.
(N) The 95th percentile box and whisker plot, number of long filopodia (>1.5 μm) emerging from the cell body (actin, n = 11; DiaΔDAD, n = 30; Ena, n = 31; DiaΔDAD+Ena, n = 16).
(O) Filopodia lifetimes (actin, n = 34; DiaΔDAD, n = 31; Ena, n = 33; DiaΔDAD+Ena, n = 14).