Supplementary Figure 10: Local injection of muscimol into M1 impairs urination; and histological confirmation of ablation of M1 neurons in the area corresponding to the region identified by PRV.

a, Schematics of the experimental protocol. b, Left, schematics showing the injection sites. Right, post-hoc histology showing the injection site. c, Quantification of the effects of muscimol or ACSF injection into different brain regions on urination(muscimol into M1: 12 male mice; ACSF into M1: 10 male mice; muscimol into Ma: 10 male mice; muscimol into the PMC: 10 male mice; ACSF into the PMC: 10 male mice; one-sided Wilcoxon signed-rank test for muscimol group compared with washout group or one-sided Wilcoxon rank-sum test for the other comparisons (***P = 0.000034, 0.000031, 0.000073, **P = 0.0012, 0.0028; z = −3.039, −3.986, −4.004, −2.772, −3.796; df = 11, 20, 20, 9, 18 from left to right). d, Quantification of the effects of muscimol or ACSF injection into the M1 of female mice on urination (muscimol into M1: 7 female mice; ACSF into M1: 7 female mice). One-sided Wilcoxon rank-sum test, ***P = 0.0008 (z = −3.154; df = 12). e, Serial coronal sections showing the NeuN staining in red and DAPI in blue in the M1 area 4 weeks after co-injection of AAV-CaMKII-Cre-GFP and AAV-DIO-taCasp3. This experiment was repeated independently in 11 male mice with similar results. All bar graphs and error bars represent mean ± s.e.m.