Fig. 1: Imaging key DCV proteins on single vesicles with light and electron microscopy.

a TIRF microscopy images of PC12 cells co-transfected with NPY-mNG (first row), Rab27a-mCherry (second row), and their overlay (third row). Left column shows representative cell image with scale bar, 3 μm. Middle column shows the enlarged images from white boxes on left column figures. Scale bar is 0.5 μm. Small regions from 5 cells normalized to the brightest pixel and averaged together (right column, scale bar is 0.5 μm). b Correlation analysis of 8 proteins with NPY-mNG–labeled DCVs in unroofed PC12 cells. Left most bar labeled NPY is a control of cells co-transfected with NPY-mNG and NPY-mCherry. “n” next to the proteins denotes the number of cells analyzed from one correlative experiment. Pink boxes are the 25th–75th percentile range of data, and the whiskers are the SD. The solid bar is the median, and the small dash is the mean. The × marks above and below each data set are the 1st and 99th percentiles. c TIRF NPY-mNG image (cyan) overlaid with PREM image (gray) of the unroofed PC12 cell membrane. This image was selected from one of the four independent experiments performed for Rab27a immunolabeled PC12 cells. Single DCV and DCV cluster (right panel) from white boxes in the left panel. Honeycomb vesicle in the top panel is a clathrin-coated pit. Scale bars are 1 μm and 100 nm for left and right panels, respectively.