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Additional Supporting Information may be found in the online version of this article.

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CNE_22679_sm_suppfig1.tif4792KSupporting Figure 1 Assessment of myenteric neuron proportions (magenta-green depiction of text Figure 1). Relative proportions of myenteric neuron phenotypes were assessed using double-label immunofluorescence or combined immunohistochemistry/immunofluorescence. A. Myenteric ganglion from jejunum demonstrating double-label immunofluorescence for NOS and HuC/D. Double-labeled NOS-positive neurons (arrows) are white. NOS-negative neurons (HuC/D only) are magenta. NOS-positive processes are green. B. Myenteric ganglion from jejunum demonstrating ChAT immunohistochemistry. C. Fluorescent image of the same ganglion shown in B demonstrating quenched HuC/D fluorescence in the ChAT-positive neuron (arrow) and a ChAT-negative (HuC/D only) neuron in magenta. Scale bar = 20 μm.
CNE_22679_sm_suppfig2.tif3623KSupporting Figure 2 Co-expression of neurotransmitter markers in myenteric plexus in Rhesus monkey (magenta-green depiction of text Figure 5). A. Co-labeling for TH (magenta) and NOS (green) revealed minimal overlap in all GI segments examined. Section depicted is from duodenum. B. Co-labeling for TH (magenta) and VIP (green) revealed minimal overlap in all GI segments examined. Section depicted is from colon. C. Co-labeling for NOS (magenta) and VIP (green) revealed significant overlap (white) that varied by segment. Section depicted is from duodenum. Overlap in circular muscle processes is also prominent. Scale bar = 20 μm.
CNE_22679_sm_suppfig3.tif2450KSupporting Figure 3 Immunohistochemical labeling of a TH-positive neuron in monkey ileum myenteric plexus. TH-positive neuron identified by brown reaction product. TH-negative neurons (HuC/D only) are magenta.

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