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. 2015 May 5;290(25):15475–15486. doi: 10.1074/jbc.M115.636670

FIGURE 5.

FIGURE 5.

In vivo FLIM-FRET analysis of interactions between Mgats and UDP-sugar transporters. A–D, mean GFP lifetime values in the absence and in the presence of acceptor are shown. The data are shown as means ± S.D. from several measurements of the indicated cell number (n). Statistically significant (one-way analysis of variance test, p < 0.001) reduction of GFP lifetime upon co-expression of fluorophore-tagged Mgats and NGT, as well as UGT1, was demonstrated by comparing with eGFP-Mgats alone. No difference in GFP lifetime was observed when mRFP-UGT2 and eGFP-Mgat1, eGFP-Mgat2, and eGFP-Mgat5 were co-expressed. Only upon simultaneous expression of mRFP-UGT2 and eGFP-Mgat4B was a statistically significant decrease in GFP lifetime observed. E, mean FRET efficiency calculated for the FRET-positive Mgat-UDP-sugar transporter combinations. The data are shown as means ± S.D. from several measurements of the indicated (A–D) cell number (n). Mean FRET efficiency was calculated to estimate relative distances between interacting proteins (the higher the mean FRET efficiency, the shorter the distance between two proteins).