Fig. 3.

Deletion of the intronic enhancer leads to an age-dependent high-bone-mass phenotype. a Sequence similarity analysis of the intronic enhancer among human, mouse, cow and dog genomes using the ECR Browser⁵⁹ (https://ecrbrowser.dcode.org) with 90% ECR similarity across a 300-base-pair ECR length. b H3K27ac ChIP-seq profile in murine osteocytic cells (GSE54784).³⁶ The nucleic acid sequence of the intronic enhancer in mice is shown in the lower panel. The sequence region colored pink denotes the deletion region obtained with the CRISPR/Cas9 method. c Representative macroscopic images of more than three wild-type (WT) and intronic enhancer-knockout (IN-KO) mice (female, 12 weeks). d, e Representative micro-CT images (d) and micro-CT parameters (e) of the femur in WT and IN-KO mice at the age of 12 weeks (n = 6 male WT and n = 6 male IN-KO mice; n = 7 female WT and n = 4 female IN-KO mice). f, g Representative micro-CT images (f) and micro-CT parameters (g) of the femur in WT and IN-KO mice at the age of 3 weeks (n = 5 female WT and n = 9 female IN-KO mice). Micro-CT scale bars: 1 mm. The data are expressed as the mean ± SEM. P values were determined by two-way ANOVA followed by Tukey’s post hoc test (e) and two-tailed t test (g)