Supplementary Figure 3: Selective deletion of melanocortin 4 receptors (Mc4r) in pre-ganglionic cholinergic neurons in the parasympathetic nervous system (PNS) causes gluco-regulatory abnormalities.

(a) 5 h fasted blood glucose, (b) plasma insulin, and (c) 5 h fasted plasma glucagon in chow-fed mice with intact Mc4r signaling (Mc4r^flox/flox) and selective deletion in the PNS (n = 8 per genotype). * indicates p<0.05 versus Mc4r^flox/flox mice using two-tailed unpaired Student's t-tests. 18 h-fasted (t = 0 min) and glucose-stimulated changes in (d) blood glucose and (e) plasma insulin during an IPGTT (1.5 mg/kg) in Mc4r^flox/flox, Mc4r^flox/flox x Chat-cre, and Mc4r-null mice (n = 8, 8, and 7, respectively). * and ** indicate p<0.05 or 0.01 versus Mc4r^flox/flox and † indicates p<0.05 versus Mc4r^flox/flox x Chat-cre using one-way ANOVA followed by Tukey's post-hoc analyses. Similar values are shown in (f) and (g) for Mc4r^flox/flox and Mc4r^flox/flox x Phox2B-cre mice (n = 7 and 8, respectively). * indicates p<0.05 versus Mc4r^flox/flox using two-tailed paired Student's t-tests. θ indicates p<0.05 comparing basal versus clamp within genotype using two-tailed paired Student's t-tests. 5 h-fasted (t = 0 min) and insulin-stimulated (h) blood glucose and (i) plasma insulin during a hyperinsulinemic-euglycemic clamps in Mc4r^flox/flox, Mc4r^flox/flox x Chat-cre, and Mc4r-null mice (n = 8, 8, and 7, respectively). * and ** indicate p<0.05 or 0.01 versus Mc4r^flox/flox and † indicates p<0.05 versus Mc4r^flox/flox x Chat-cre using one-way ANOVA followed by Tukey's post-hoc analyses. θ indicates p<0.05 comparing basal versus clamp within genotype using two-tailed paired Student's t-tests. Similar values are shown in (j) and (k) for Mc4r^flox/flox and Mc4r^flox/flox x Phox2B-cre mice (n = 7 and 6,respectively).