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. 2016 May 5;6:25502. doi: 10.1038/srep25502

Figure 3. Development of RhoB/C- and localization of RhoA/B/C FRET sensors.

Figure 3

(a) Intramolecular RhoGTPase FRET sensor design that consists of (from N- to C-terminal) the RBD of PKN1 (ΔPKN), YFP (mVenus), CFP (Cerulean3) and a RhoGTPase. RhoGTPase activation results in RBD-RhoGTP binding and an energy transfer from CFP to YFP. (b) Average basal YFP/CFP ratios (±SEM) of RhoA-NB (n = 13), RhoA-WT (n = 16), RhoA-Q63L (n = 13), RhoB-NB (n = 17), RhoB-WT (n = 20), RhoB-G14V (n = 18), RhoC-NB (n = 14), RhoC-WT (n = 20), RhoC-Q63L (n = 14) and RhoC-G14V (n = 14) FRET sensors expressed in EC. (c) EC were transfected with RhoA/B/C-WT FRET sensors and stained for actin and VE-cadherin. Boxes show co-localization of the FRET sensors with VE-cadherin, corresponding profile plots show gray values for RhoA/B/C-WT FRET sensors (green line) and VE-cadherin (blue line) according to the lines selected in the box regions. Arrowheads indicate vesicular localization of the RhoB-WT sensor. Bar = 14 μm.