Characterization of Fbp17. (A) Sequence of the C-terminus of GxcM (aa 791–1,145). Proline residues are highlighted in red. The blue underlines indicate SH3 domain-binding motifs predicted by the LMDIPred web server. (B) Sequence alignment of Fbp17 with human CIP4, FBP17, and TOCA-1. Magenta, green, and blue boxes indicate the F-BAR, homology region 1 (HR1), and SH3 domain, respectively. (C and D) Time-lapse imaging of GFP-Fbp17 in randomly migrating cells (C) or cells moving under agarose along a folic acid gradient (D). Scale bars, 5 μm. (E) Top: Design of the fbp17 knockout construct. Bottom: Targeted clones were confirmed by PCR. (F) The indicated cells were plated clonally with bacteria (Klebsiella aerogenes) on standard medium agar for 5 d. Scale bar, 5 mm. (G and H) Left: Localization of GFP-myosin II in randomly migrating cells (G) or cells migrating under 2% agarose along a folic acid gradient (H). Right: Quantification of the relative cortical localization of GFP-myosin II. Data were from three independent experiments; the scatter plots show data points with mean ± SEM; n, number of cells analyzed. Scale bars, 5 μm.