Fig. 6

Comparative imaging of calcified deposits in mdx and control mice muscles. Triceps brachii, quadriceps femoris, and spinalis pars lumborum muscles were dissected from perfusion-based CUBIC-cleared mdx model of DMD and control mouse, stained with alizarin red S and imaged in bright-field (a1), crossed polarized light (a2), and fluorescence (a3). Arrowheads indicate calcified deposits. b Three-dimensional light-sheet reconstructions of calcified deposits in muscles of mdx mouse. c Quantitative volume analysis of frequency distribution (left), relationship between cumulative volume and counts of calcified deposits (middle), cumulative frequency distribution (right) in triceps brachii (blue line and squares), quadriceps femoris (red line and triangles), and spinalis pars lumborum (black line and triangles). Two-sample Kolmogorov-Smirnov test, *p < 0.001 quadriceps femoris vs triceps brachii, quadriceps femoris vs spinalis pars lumborum. d Comparative volume analysis of calcified deposits in muscles of mdx mouse normalized to quadriceps femoris. Number of calcified deposits in mdx mouse shown in b, c, d: triceps brachii (n = 89), quadriceps femoris (n = 652), spinalis pars lumborum (n = 548)