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. Author manuscript; available in PMC: 2019 Feb 1.
Published in final edited form as: Bone. 2017 Dec 5;107:172–180. doi: 10.1016/j.bone.2017.12.001

Figure 1.

Figure 1

LARG is required for mechanical activation of RhoA in MSCs. (A) Mechanical treatment of mdMSCs (2% strain, 100 cycles) significantly (p<0.05, n=4) increased RhoA activation (RhoA-GTP). siRNA knockdown (20 nM) of LARG prevented strain-induced RhoA activation. (B) Knockdown of Vav2 did not influence strain-induced activation of RhoA, where mechanical strain significantly increased RhoA activity in the presence (p<0.05, n=3) and absence (p<0.01, n=3) of Vav2. (C–F) Staining of the actin cytoskeleton using alexa 488-conjugated phalloidin showed increased stress fiber formation in strained cells (D) compared to static controls (C). siRNA knockdown of LARG, reduced the stress fiber formation following mechanical strain (F). (G) Quantification of stress fibers confirmed that strain significantly (p<0.0001) increased the number of stress fibers, whereas no significant increase was observed following knockdown of LARG. For actin quantification, at least 15 cells per condition total were analyzed across 3 independent experiments.