Fig 1: Immunohistochemistry and ISH analysis and quantification of overlapping PACAP- and VGLUT2-expressing cells in the BMA. A, Representative photomicrographs from tissue sections containing the BMA from PACAP-EGFP mice represent expression of VGLUT2 (red) and PACAP-EGFP (green) processed with immunohistochemistry for GFP and VGLUT2. White arrows indicate overlapping VGLUT2 and PACAP signals. Green arrow indicates an example of PACAP cell without VGLUT2 expression. Scale bar, 25 µm. Right, Quantification of overlap (N = 6: 3 M and 3 F; df = 5). B, Representative photomicrographs from tissue sections containing the BMA from WT mice represent overlapping VLGUT2 (green) and PACAP mRNA (red) fluorescence in the BMA. Tissues were processed with RNAscope technique for PACAP mRNA and then immunohistochemistry for VGLUT2. Right, Venn diagram represents quantification of overlap between VGLUT2 and PACAP mRNA (N = 5: 2 M and 3 F; df = 4).