Fig. 2.

Distribution of NiV envelope glycoproteins on the plasma membrane of PK13 cells. PK13 cells expressing NiV-F or/and -G were fixed at 24 h post transfection and immunostained. Without permeabilization, NiV-F was immunostained by using a mouse anti-FLAG primary antibody and -G a rabbit anti-HA primary antibody. For single-color SMLM, Alexa Fluor 647 secondary antibodies were used for detection. For dual-color SMLM, Alexa Fluor 647 secondary antibodies were used for detection of F, and Cy3B secondary antibodies for G. a, b x–y cross section (100 nm thick in z) at the dorsal surface (position 3 in Fig. 1a) of a representative cell expressing F (a) or G (b). The boxed region is enlarged to show the detailed distribution pattern. c Comparison analyses of the localization densities of the F (red) or G (green) at the cell body versus membrane protrusions. Each data point was calculated using an area of 0.2 × 0.2 μm². All data were normalized to the mean of the cell body. d Hopkins’ index of the F and G localizations from n = 30 cells. Lines represent the mean value and SD. The sample size is indicated in the parentheses. The p values were determined by two-tailed, unpaired t-test with Welch correction. e x–y cross section (100 nm thick in z) of a region at the dorsal surface (position 3 in Fig. 1a) of a representative cell co-expressing F (red) and G (green) with a pixel size of 10 nm. Scale bars: 1 μm. f The distribution of the DoC values between F and G molecules. One representative cell image out of three independent experiments (n ≥ 30) is shown