Fig. 5: Nanostructure guided CHIKV replication.

a SEM image of gradient nanorings with decreasing inner ring diameters, ranging from 1500 nm to 700 nm. n = 3 repeats. b Schematic of a nanoring illustrating three distinct positive curvatures on PM: (i) the sidewall of the outer ring; (ii) the top rim of the outer ring; and (iii) the top rim of the inner ring. c Confocal images of CHIKV-infected cells cultured on a flat surface or a nanoring array. Replication sites were indicated by dsRNA staining (magenta). n = 3 repeats. d Confocal images of nsP3-mCherry tagged CHIKV-infected cells cultured on the nanoring array. The mCherry fluorescent tag was inserted at the hypervariable region of nsP3 (as shown in cyan), which is known to tolerate tags and does not impede virus replication and infection. The cells were stained for nsP1 (yellow) and dsRNA (magenta). n = 3 repeats. e Comparison of the dsRNA density in the inner ring of the nanoring area and on the nearby flat surface (n = 60 rings). f Average images of the brightfield, PM intensity, and dsRNA intensity on different-sized nanorings (over 30 rings for each size). g The dsRNA intensity per unit inner rim area on different-sized nanorings (n = 27, 31, 44, 60, 39). Mean ± SD. h The dsRNA intensity per unit inner rim perimeter on different-sized nanorings (n = 27, 31, 44, 60, 39). Mean ± SD. Statistical analysis: two-tailed unpaired t-test. Scale bars: 2 μm. Arb. units = arbitrary units. Source data are provided as a Source Data file.