Fig. 1: Single-molecule fluorescence assay of SNARE-nanodisc containing Vamp full, SN25 and Stx-Cy3 by NSF. | Nature Communications

Fig. 1: Single-molecule fluorescence assay of SNARE-nanodisc containing Vamp full, SN25 and Stx-Cy3 by NSF.

From: Single-molecule two- and three-colour FRET studies reveal a transition state in SNARE disassembly by NSF

Fig. 1: Single-molecule fluorescence assay of SNARE-nanodisc containing Vamp full, SN25 and Stx-Cy3 by NSF.

a Procedure of the single-molecule SNARE disassembly assay on total internal reflection fluorescence (TIRF) microscope. b TIRFM images of Cy3 fluorescence channel containing SNARE-nanodisc after immobilisation along with a pictorial representation of nanodisc present on the glass slide before and after the disassembly by NSF. c Number of Stx-Cy3 nanodiscs present under various experimental conditions. Disassembly reaction in the presence of all components (brown), without the addition of Mg2+ (light grey), only with ATP already bound to the NSF (No extra ATP) and other controls. p-values calculated using a two-tailed paired t test and mean ± SD values are given, ***= p < 0.005. (n = 3-6 independent SNARE disassembly experiments).

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