Fig. 4.

GAS7b sheets on liposomes and phagocytic cups of macrophages. a Illustration of the possible spatial distributions of super-resolution signals. Randomly distributed signals, signals from randomly placed GAS7b dimers and signals from GAS7b forming FFO sheets are schematically illustrated. The alignment of the FFOs in the simulated sheet was considered to be lateral, as illustrated. Dashed magenta circles indicate the counting of the signals neighbouring a signal. b–e Phase-contrast (b, d) and epi-fluorescent (c, e) images of liposomes (b, c) and phagocytic cups (d, e) using mEOS4b-GAS7b, with arrows indicating incorporated zymosan. Scale bar: 5 μm. f–i Super-resolution images showing the single-molecule localization of photoconvertible mEOS4b-tagged GAS7b. Left panels are the 60 nm slices of mEOS4b signals on glass (f), on liposomes (g), at the bottom of a phagocytic cup (h), at the top of a phagocytic cup (i) and at the indicated observation depths of the regions marked by red rectangles in b, c and d, e. Middle panels are enlarged images of the parts enclosed in red rectangles in the left panels. Graphs on the right show the occurrences of observed neighbour signals (shown in magenta) dependent on the neighbour distances within the regions enclosed by red rectangles in the left panels. The same numbers of signals from the simulated randomly placed GAS7b dimers and the simulated FFO sheet, under the assumption of the 10% observation of the total molecules, are shown in green and blue, respectively. The occurrence is described as the fold increase in the same numbers of the randomly distributed simulated signals. Dashed lines represent ±SD of 20 trial simulations. Scale bars: 1 μm (left) and 100 nm (middle)