Actin stabilization mediated by AnxB9 is required for RhoGEF2 recruitment to cell wounds. (A–C) Actin dynamics during cell wound repair in control (GAL4 driver 7063 alone) (A), AnxB9^RNAi(1) (B), and AnxB9^RNAi(2) (C) NC4–6 staged embryos expressing an actin reporter (sGMCA). (A′–C′) xy Kymograph across the wound area in A–C, respectively. Arrowheads denote actin accumulation within the wound. (D) Quantification of the wound area over time (control, AnxB9^RNAi(1), and AnxB9^RNAi(2); n = 10 respectively). Error bars represent ± SEM (E–G″) Confocal xy projection images from Drosophila NC4–6 staged embryos coexpressing sfGFP-RhoGEF2 and sChMCA in an AnxB9^RNAi background injected with buffer (E–E″) or phalloidin (F–G″). RhoGEF2 partially accumulates at the wounds (arrowheads; F). (H) Schematic diagram summarizing the localization patterns of actin, AnxB9, RhoGEF2, Pbl, RhoGEF3, Tum, Rho1, Rac1, and Cdc42 at cell wounds. (I) Schematic diagram summarizing the pathways among AnxB9, actin, RhoGEFs, Tum, and Rho family GTPases in response to cell wounds. Time after wounding is indicated. Bars, 20 µm.