Fig. 2
Msn is required for BC migration, protrusion restriction and detachment. a Quantification of cluster velocity of control (n = 10) and Msn-depleted clusters (n = 8) from live imaging. ***p < 0.001, unpaired Student’s t-test. The error bars show s.e.m. Quantification of the mean square displacement (MSD) over time in b and cumulative in c of control (n = 10) and Msn-depleted clusters (n = 8) from live imaging. ***p < 0.001, two ways ANOVA test. On box plots, the box limits represent the first (lower) and the third (upper) quartile values, the whiskers show the smallest and the greatest values; and the center line represents the median. n represents the numbers of independent BC clusters. d Inverted still greyscale images from a time-lapse recording of clusters expressing LifeAct::GFP in control or Msn-depleted conditions. Black arrows indicate protrusions and red arrows point to the detachment defect (see Movie 2). e Using 2D live imaging, we quantify the number of protrusions per cluster at each time point of 5 h movies of control and Msn-depleted clusters (n = 8 and 10, respectively). Any protrusions larger than 4 μm were automatically detected using ADAPT plugin and LifeAct::GFP as a marker. **p < 0.01, two ways ANOVA test, The error bars show s.e.m. f Quantification of the percentage of clusters that started to migrate and still have not detached by stage 10 (fixed samples) (n = 47 and 39, for Control and msn RNAi respectively)
