Figure 5

NET labeling segregates from markers of large and small dense core vesicles. SCGs were double-labeled with NET-05 and antibodies directed against VMAT2, NPY or synaptophysin. The first column (A, E, and I) represents the DIC images of an exemplary field of cells, whereas the second column (B, F, J, green) is the distribution of NET in the same or fibers or boutons. The fourth column (C, G, K, red) are images obtained with antibodies against NPY, VMAT2, or synaptophysin (Synapto). The last column (D, H, and L) represents the overlap of the signals obtained with NET-05 and the particular granule protein targeted. Scale bar = 10 micron. Insets in staining panels provide higher power views of single boutons. Scale bar = 5 micron. M. Correlation of NET and VMAT2 pixel densities (upper left), ICA analysis (lower panels), and spatial display of PDM values (upper right). N. Correlation of NET and synaptophysin pixel densities (upper left), ICA analysis (lower panels), and spatial display of PDM values (upper right). PDM color transformation is shown above standard red/green visualizations of same field.