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. 2011 May 3;286(26):23308–23318. doi: 10.1074/jbc.M111.217950

FIGURE 4.

FIGURE 4.

NBF1 and NBF2 truncated proteins containing Walker domains can disrupt Syn-1A interaction with SUR1 in live cells. A, NBF1 and NBF2 can inhibit Syn-1A interaction with SUR1. Panel (i), representative recording of FRET signals on the plasma membrane of HEK cell expressing Syn-1A-mCherry, SUR1-EGFP, and Kir6.2 before and after addition of 1 μm truncated NBF1/NBF2 proteins to the permeabilized cells in the reverse sequences (NBF1 then NBF2; or NBF2 then NBF1). The excitation wavelength was 488 nm, and fluorescence emission was measured at 605–655 nm. The scale bar indicates 5 μm. The vertical scale bar indicates FRET efficiency in pseudocolor. Panel (ii), summary of FRET efficiency before and after addition of different NBF truncations. The bar graph shows the means ± S.E., n = 15. ***, p < 0.001. B, WB but not WA domain of NBF1 can inhibit the interaction between Syn-1A and SUR1. Panel (i), representative recordings of the FRET signal on the plasma membrane of one same HEK cell before and after addition of 1 μm NBF1 truncations in different sequence (WA, linker then WB; or WB, linker then WA). Panel (ii), bar graph (means ± S.E.), n = 5. ***, p < 0.001. C, both WA and WB domains of NBF2 can inhibit the interaction between Syn-1A and SUR1. Panel (i), representative recordings of the FRET efficiency on the plasma membrane of one same HEK cell before and after addition of 1 μm NBF2 truncations in different sequence (WA, linker, and then WB; or WB, linker, and then WA; or linker, WA, and then WB). Panel (ii), corresponding bar graph (means ± S.E.), n = 7. ***, p < 0.001; *, p < 0.05.