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. 2011 Dec 12;108(52):21010-21015. doi: 10.1073/pnas.1113651108

Fig. 2.

Fig. 2.

MDA5 forms filamentous oligomers along the length of dsRNA. (A) Representative electron micrographs of negatively stained MDA5 in complex with 112 and 512 bp dsRNA in the presence of ADP•AlF4. Representative particles used for averaging are circled in red. See Fig. S2C for electron micrographs of MDA5 in complex with ssRNA and dsDNA. (B) Representative class averages of MDA5 in complex with 112 bp dsRNA. The red box in the magnified view corresponds to a minimum binding unit of MDA5 in EMSA (C). The number of binding units per complex (nb) is displayed at the bottom left for each average. (See also Fig. S2D.) (C) EMSA of 62, 112, and 162 bp dsRNAs with MDA5 (0.3 μM) visualized with SybrGold stain to enhace the intermediate complexes that are otherwise difficult to detect. Enumeration of intermediate complexes reveals that the full complex contains 4, 7–8, and 11 binding units of MDA5 for 62, 112, and 162 bp dsRNAs, respectively. (D) Multiangle light scattering (-OD280 and ▪ Mr estimation) suggests that the molecular mass of the full complexes formed on 62 and 112 bp dsRNAs are 854 ( ± 43) kDa and 1,878 ( ± 13) kDa, respectively.