Figure 5.

INSL5, PYY, and GLP-1 are typically localised within the same vesicles in murine colonic cultures. (A) 3D-SIM image of murine colonic culture labelled for INSL5, PYY, and GLP-1. Images are projected in $z$. Scale bar = 5 μm. Insets represent individual $z$-planes of vesicle clusters, scale bar = 500 nm. (B) Scatter of normalised intensity in PYY (top) and GLP-1 (bottom) relative to INSL5 (n = 9860 vesicles, 17 cells, 4 exp.). Dashed lines represent the intensities above (+, 5% of maximum) and below (–, 2%) which vesicles were considered positive or negative, respectively, for each peptide. (C) Percentage of vesicles per cell that contain single peptides (brown), two peptides (blue; double), or all three (red; triple). IDs represent the experiment (e) and cell (c) numbers, in the format e.cc. The final line (‘All’) represents the percentages of vesicles across all experiments. (D) Example vesicle signals in each INSL5, PYY and GLP-1 categorised by whether the vesicle contains only a single peptide (first three rows), pairs of peptides (next three rows), or all three peptides (bottom row). Percentages represent percentage of vesicles within each category. (E) Mean intensity profiles through vesicle signals for each category in D for INSL5 (green), PYY (red) and GLP-1 (blue). Shaded regions represent standard deviation.