Myo1b KD reduces the number of large islets formed by YFP-EphB2-HCT116 cells when they are cocultivated with Cherry-ephrinB1-HCT116 cells. (A and B) YFP-EphB2-HCT116 cells transfected with control or Myo1b siRNAs and cocultivated with Cherry-ephrinB1-HCT116 or Cherry-HCT116 cells were analyzed by phase-contrast and fluorescent microscopy. (A) The overlay of phase-contrast images (gray) and YFP images (green). (B) The same images as A after treatment as described in Fig. S3 to visualize the outlines of the islets formed by YFP-EphB2-HCT116 cells. Bar, 150 µm. (C) The number of cells per islets was quantified in the different experimental conditions based on the nuclear staining and as described in Fig. S3. Data are shown as the mean of three experiments. Error bars represent ± SEM. n = 191 and 346 islets for control siRNA-treated YFP-EphB2-HCT116 cells cocultivated with Cherry-HCT116 cells or with Cherry-ephrinB1-HCT116 cells, respectively. n = 198 islets for Myo1b siRNA-treated YFP-EphB2-HCT116 cells cocultivated with Cherry-ephrinB1-HCT116 cells. Paired Student’s t test was used to analyze the probabilities of these data. ***, P = 7.7 × 10−7 for control siRNA YFP-EphB2-HCT116 cells cocultivated with Cherry-HCT116 cells versus those cells cocultivated with Cherry-ephrinB1-HCT116. ***, P = 1.7 × 10−8 for Myo1b siRNA YFP-EphB2-HCT116 cells cocultivated with Cherry-ephrinB1-HCT116 cells versus control siRNA YFP-EphB2-HCT116 cells.
