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. Author manuscript; available in PMC: 2017 Apr 28.
Published in final edited form as: Methods. 2016 Mar 25;105:99–108. doi: 10.1016/j.ymeth.2016.03.022

Fig. 3.

Fig. 3

Single-molecule stoichiometry measurement. A. Schematic overview of a single-molecule stoichiometry measurement. Biotinylated Drosha-DGCR8/eGFP complexes were immobilized on a PEGylated surface using biotin-streptavidin conjugation. Cy5-labeled pri-miRNA molecules were introduced into the imaging chamber. B. Representative time trajectory of eGFP fluorescence. The stoichiometry of DGCR8 molecules associated with Drosha molecules was determined by counting the number of eGFP photobleaching steps (indicated with dashed lines). C. Bar plot showing the distribution of the Drosha/DGCR8 stoichiometry. D. Ability of Drosha-DGCR8 complexes to bind RNA molecules was confirmed using the co-localization of pri-miRNA and DGCR8. The figure shows a camera screenshot with eGFP molecules in the left channel and Cy5 molecules in the right channel. The red circles indicate co-localized DGCR8 (eGFP) and pri-miRNA (Cy5).