Figure 1: aVSOP reveals an association between functional bladder capacity and Cx43.

(a) A photograph and diagram showing the aVSOP method. Each stain was traced, scanned and quantified by Image J 1.42 software. (b–d) Female Cx43^+/− mice had larger functional bladder capacity than sex-matched Cx43^+/+ littermates. (b) A photograph of urine spots on paper made by Cx43^+/+ (top) and Cx43^+/− (bottom) mice. The scale bar indicates 10 cm, corresponding to 1 h. (c) Representative charts of UVVM of Cx43^+/+ (top) and Cx43^+/− (bottom) mice under LD conditions for 4 days. (d) UVVM per 6 h in Cx43^+/+ and Cx43^+/− mice had diurnal variation (F(3[degrees of freedom (DF) for the time factor],9[error DF])=12.3 and 10.9, respectively; *P<0.005 by one-way repeated measures ANOVA; P<0.05 in the late light (sleep) phase versus late dark (active) phase, followed by Bonferroni's post hoc test). Maximal correlations from a cosine curve (MaxCorr) of Cx43^+/+ and Cx43^+/− mice were 0.949 and 0.989, respectively: light-on at ZT0 and off at ZT12. UVVM was significantly different between Cx43^+/+ and Cx43^+/− mice (F(1[DF for the strain factor],6[error DF])=11.2, P<0.05 by two-way repeated measures ANOVA; †P<0.05 versus Cx43^+/+ by Bonferroni's post hoc test; n=4 for each group, with a total of 296 micturitions). Error bars represent s.e.m. (e) Relative Cx43 mRNA levels of the urinary bladder in Cx43^+/− and Cx43^+/+ mice used in the micturition analysis by real-time RT–PCR. Error bars represent s.d., n=4 for each mice. The value of Cx43^+/+ was set as 1. *P<0.05 by Student's t-test. (f) Cx43 protein expression of the urinary bladder in Cx43^+/+ and Cx43^+/− mice.