Supplementary Figure 4: Fingers are under RhoA control.
From: Interplay of RhoA and mechanical forces in collective cell migration driven by leader cells
(A) Use of chemical inhibitors. The drugs were used either at the beginning of the experiments or 4–6 hours after stencil removal to highlight their action on finger formation or on their maintenance. Leaders are identified by their phenotype (larger cells, active ruffling lamellipodium). Bars are 250 m. n = 6 independent experiments. Errors are SEMs. (B) Validation of anti-RhoA siRNAs. Western blots anti-Rac1 and anti-RhoA were performed on lysates of MDCK cells not-transfected or treated with control siRNA, siRhoA#1 or siRhoA#2. Expression levels were normalized towards GAPDH (not shown) and the no-transfection condition was used as reference. (C) Distribution of RhoA differential activity along a finger in presence of C3 Transferase. The Rho inhibitor results in a strong decrease of Rho (Blue). This decrease mirrors the response of the force profile in similar RhoA-inhibition conditions (Fig. 4B). Red points are the RhoA differential activity with no RhoA inhibition (from Fig. 5F). Observations were performed after 2 h incubation of C3 transferase on n = 16 cells spanning 12 well-defined fingers from 2 independent experiments. Error bars are SEMs.
