Fig. 1
Experimental overview and microcomputed tomography (μCT)-based quantification of ectopic calcification in ENT1−/− mice. A Wild-type (10 female, 10 male) and ENT1−/−mice (10 female, 10 male) were studied for behavioral indicators of pain and physical function at 2, 4, and 6 months of age. Behavioral assays were conducted over three testing days. µCT was performed to correlate the extent of ectopic calcification with behavior. B Three-dimensional renderings (anterior view) of the same representative male ENT1−/− mouse at 2.5, 4.5, and 6.5 months of age. Inserts present magnified posterior view of the cervical-thoracic spine. Areas of hyperdense tissue, exceeding the radiodensity of vertebral cortical bone (≥ 1278 HU), are pseudocolored in red. C Ectopic calcification in ENT1−/− mice quantified as the ratio of hyperdense tissue to vertebral bone (C1 to T12). Linear regression analysis with goodness of fit values presented as r2. Each animal is represented as a unique symbol. The slopes of the linear regressions were significantly non-zero with no differences between sexes (female, y = 0.0093x – 0.0007; male, y = 0.0096x – 0.0053). Data were pooled to analyze the overall relationship between ectopic calcification and age using the mean value for each time point (y = 0.0094x – 0.0030). Red lines represent the linear regression with dotted lines representing 95% confidence intervals