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. 2017 Nov 7;28(23):3371–3382. doi: 10.1091/mbc.E17-03-0136

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© 2017 Reinhard et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 7: — Gα₁₃ inhibition enhances S1P-induced Rac1 activation and blocks RhoA activation. (A) Normalized mean YFP/CFP ratio traces (±SEM) for Rac1, Cdc42, and RhoA FRET sensor-expressing ECs before and after stimulation with S1P at t = 1 min, 50 s. FRET sensor–positive ECs coexpress either Lck-mCherry (control) (Rac1, n = 20; Cdc42, n = 21; RhoA, n = 14) or Lck-RGS-mCherry (RGS) (Rac1, n = 27; Cdc42, n = 22; RhoA, n = 14). (B) Enlargement of normalized mean YFP/CFP ratio traces (±SEM) corresponding to the first 7 min of A. Dashed lines highlight maximal and minimal YFP/CFP ratio changes. Note the increased Rac1 activation at ∼7 min and the inhibition of RhoA activation upon coexpression of the RGS domain. (C) Model describing S1P-mediated signaling pathways in ECs. Balancing G-protein and RhoGTPase signaling results in S1P-induced cell spreading, which dominates over the limited RhoA activation and consequent contraction.