FIGURE 7:

LARG and GEF-H1 knockdowns decrease cell stiffness and stiffness response to force and increase cell migration and invasion. (A) NMuMG cells were transfected with siRNA against LARG, GEF-H1, both, or p114 for 48 h. Quantifications are given normalized to β-actin and relative to control, confirming knockdown of protein expression. (B) Average cell stiffness (G₁) for NMuMG cells treated with siRNA control (n = 88), siRNA targeting p114 (n = 70), GEF-H1 (n = 90), LARG (n = 85), GEF-H1 + LARG (n = 100), and NMuMG cells treated with TGF-β and siRNA control (n = 69). *p < 0.01. Error bars represent SEM; data were collected from three independent experiments. (C) Average stiffness response at pulls 2 (G₂/G₁) and 8 (G₈/G₁) for NMuMG cells treated with siRNA control (n = 20), siRNA targeting p114 (n = 41), GEF-H1 (n = 19), LARG (n = 21), GEF-H1 + LARG (n = 30), and NMuMG cells treated with TGF-β and siRNA control (n = 19). ^#p < 0.05 stiffness difference from G₁; *p < 0.05 stiffness response difference relative to cells treated with siRNA control. Error bars represent SEM; data were collected from three independent experiments. (D, E) Average migration and invasion for NMuMG cells treated with siRNA as indicated: control, GEF-H1, LARG, both, or p114 for 36 h before plating onto uncoated or Matrigel-coated Transwell filters. *p < 0.05. Error bars represent SEM; data represent mean of three independent experiments.