a, Representative ECG traces obtained from sedentary, trained and detrained mice (unrestrained and conscious). b, Mean (+SEM) heart rate measured in vivo (n=7/8/5) and in vitro (from isolated sinus node preparations; n=7/8/5) in sedentary, trained and detrained mice. c, Restoration of the contribution of If to pacemaking in the detrained mouse. Mean (+SEM) percentage decrease in heart rate of isolated sinus node preparations from sedentary, trained and detrained mice on blocking If using 2 mM CsCl shown (n=6/7/4). d-f, Restoration of normal levels of HCN4 and Tbx3 mRNA and miR-1 in the sinus node in the detrained mouse. Mean (+SEM) expression of HCN4 and Tbx3 mRNA (normalised to 18S) and miR-1 (normalised to RNU1A1) in the sinus node of sedentary, trained and detrained mice shown (n=4). Student’s t test used to test differences. Normal distribution of data was tested using the Shapiro-Wilk W test and equal variance was tested using the F test. When the null hypothesis of normality and/or equal variance was rejected, the nonparametric Mann Whitney U test was used. *P<0.05, trained versus sedentary mice; †P<0.05, detrained versus trained mice.