Panel a shows representative wound-healing images for shControl, shPACSIN2, shARHGAP17, shASAP1, shBIN1, shBIN3, shFER, shPICK1, shSH3GL1, and shSNX9. White circles mark nuclei, colored tracks indicate cell migration, and the color scale represents track displacement from 1 to 120 micrometers. Scale bar = 50 micrometers. Panel b shows a bar graph of wound closure (percent) versus shRNA, comparing shControl, PACSIN2, ARHGAP17-1, ARHGAP17-2, ASAP1-2, BIN1-1, BIN1-2, BIN3-1, BIN3-2, FER-1, FER-2, FER-3, FER-4, FER-5, PICK1-1, PICK1-2, SH3GL1-1, SH3GL1-2, SNX9-1, and SNX9-2, with individual data points, error bars, a dashed reference line, and significance indicated by asterisks. Panel c shows a bar graph of percentage of Golgi oriented toward the leading edge versus shRNA for the same conditions, with individual data points, error bars, a dashed reference line, and significance indicated by asterisks.
BIN1, FER, and SNX9 control endothelial collective cell migration. (a) Representative widefield images of migrating HUVEC monolayers transduced with shRNAs targeting the indicated BAR proteins, captured 2 h after scratch wound formation. Nuclei are stained with Hoechst (greyscale). Migration tracks, shown for the 2–9 h period after scratch, illustrate differences in cell migration dynamics between conditions. Scale bar = 50 μm. (b) Quantification of endothelial sheet wound closure 9 h after scratch. For each shRNA, at least n = 3 movies were quantified from at least three independent experiments. (c) Quantification of Golgi orientation in the first four rows of follower cells at the scratch edge. For each shRNA, at least n = 5 images per experiment were quantified from at least three independent experiments. Data are presented as mean ± SD, and the statistical analysis was performed using one-way ANOVA, followed by Dunnett’s test for multiple comparisons versus shControl. * = P < 0.05, ** = P < 0.01, and *** = P < 0.001.