Fig. 5

Vangl2 regulates RhoA activity in leader cells of collectively migrating breast cancer cells. a Representative spatial activity profiles of RhoA in collectively migrating MCF7 cells stably expressing RhoA-FRET biosensor and Control (top row), shVangl2-1 (middle row), or shVangl2-2 (bottom row) at 1 h (left column), 6 h (center column), and 12 h (right column. Color bars indicate the range of RhoA-FRET biosensor ratios. Scale bar = 25 µm. b RhoA activity as a function of distance in µm from the leading edge of collectively migrating MCF7 cells stably expressing Vector (n = 27 wells), shVangl2-1 (n = 24 wells), or shVangl2-2 (n = 25 wells) at 1, 6, and 12 h of migration, error bars indicate ± sem. c, d RhoA activity after 1 h of migration at 5 µm (c) and 100 µm (d) from the leading edge of collectively migrating MCF7 cells stably expressing Vector, shVangl2-1 (Vector vs shVangl2-1, 5 µm p = 0.00388, 100 µm p = 0.1010, or shVangl2-2 (Vector vs shVangl2-2, 5 µm p = 0.0384, 100 µm p = 0.1309), significance was determined by a two-sided unpaired t test