Figure 1. RhoA enhanced cytoskeleton modulates LMS-induced FAK phosphorylation.

a) Schematic of time course experiment depicting different groups. Groups (1) and (2) were subjected to single LMS with or without a 2h rest, respectively. Group (1+2) was subjected to both LMS treatments. b) LMS acutely increased FAK phosphorylation 3.4-fold (p<0.001), 2h later p-FAK was not significantly different than control. Re-application of LMS resulted in 7-fold increase, doubling the single LMS response. c) Isolation of FAs 2hr after a single LMS treatment was subject to Western blot analysis; increased total vinculin, paxillin and Akt were consistent with increased FAs. d) FA increase was accompanied by RhoA activity and e) RhoA activity was dependent on initial LMS-induced FAK activity as FAK inhibitor PF573228 (PF, 3µM) prevented LMS-activated RhoA. f) RhoA activated with LPA (30µM) increased p-FAK 3-fold (p<0.05); following LPA, a single LMS application increased p-FAK response by 7-fold (p<0.001). The black bar on the representative western blot indicates that it has been cropped to change the sample positions. g) LMS amplified the HMS response: 1 and 2X LMS pretreatment synergistically augmented the HMS response by 30% (p<0.01) and 60% (p<0.01), respectively. * p<0.05, ¥ p<0.01, † p<0.001, against control and each other.