Videos of MDA-MB-231 LifeAct-Ruby2 or MIIA-GFP cells were analyzed by a custom MATLAB script. In short, the cortical actin and myosin signals of the protrusions within each branch channel were obtained after background subtraction of the fluorescence signal of each cell. For the normalized integrated signal intensity over a range of hydraulic resistances, all values were normalized to the signal on the 1× relative resistance branch.

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