An RNA aptamer that distinguishes between closely related human influenza viruses and inhibits haemagglutinin-mediated membrane fusion, by S. C. B. Gopinath, T. Misono, K. Kawasaki, T. Mizuno, M. Imai, T. Odagiri and P. K. R. Kumar
Journal of General Virology vol. 87, part 3, pp. 479 – 487
Supplementary Methods
Supplementary Fig. S1. SPR analysis of the P30-10-16–HA complex.
Supplementary Fig. S2. SPR analysis of the mAb–HA complex.
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Supplementary Video S1 (P30-10-16C). Inhibition of HA-mediated membrane fusion by P30-10-16C RNA. A time sequence of fluorescent images of virus–ghost conjugates during membrane fusion in the presence of complementary RNA P30-10-16C (5 µM) is demonstrated. After installing conjugates of R18-A/Panama and RBC ghosts on a fluorescence microscope at pH 7·5 and 20 ºC, a low-pH trigger (pH 5·0) was applied at time zero (the beginning of the video). The R18 fluorescence, which was originally constrained at punctate regions of individual virus particles, diffused out to the larger regions of the RBC ghosts, which indicates inter-membrane lipid mixing due to viral membrane fusion. The time sequence of 60 time-lapse images, recorded over 180 s (one frame every 3 s), is replayed with 36-fold-enhanced speed (12 frames s–1). [AVI file]
Supplementary Video S2 (P30-10-16). Inhibition of HA-mediated membrane fusion by P30-10-16 RNA. A time sequence of fluorescence images of virus–ghost conjugates after the low-pH trigger (pH 5·0 and 20 ºC, at time zero) in the presence of the selected aptamer P30-10-16 (5 µM) is demonstrated. In contrast to the result shown in Supplementary Video S1, R18 redistribution between the virus and the RBC ghost was not observed at most of the virus–ghost conjugates, indicating the inhibitory effect of the aptamer on viral membrane fusion. The images were recorded and replayed as described for Supplementary Video S1. [AVI file]
