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. 2015 Mar 12;10(3):e0116900. doi: 10.1371/journal.pone.0116900

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© 2015 Storti et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Fig 1 — Panel a-e refer to sensitized emission (steady state) FRET. (a) Donor (Caveolin-EGFP) emission image; (b) acceptor (TRPV1-RFP) emission image; (c) SE-FRET intensity image (scale bar: 5 μm); (d) zoom of a cellular region where FRET-iMSD analysis is later performed; (e) donor-normalized apparent FRET efficiency E _D. Panels f-g refer to FRET-iMSD analysis. (f) iMSD vs. time plot of FRET signal for the Caveolin-1-EGFP/TRPV1-RFP pair in physiological condition. The constant iMSD clearly demonstrates that the complex is not diffusing in space, i.e. it is trapped within caveolin-positive structures on cellular membrane. (g) Temporal evolution of the binding component: plot of G(0,0,τ) vs. time shows the decrease of correlation function in time. This decrease of STICS function associated to a constant iMSD plot (f) confirms the presence of a binding interaction between TRPV1 and caveolin-1.