Micro-computed tomography (µCT)

The skeletal phenotype of BgnFmod KO mice was first analyzed in whole femurs and L3 lumbar vertebrae using the µCT50 system (Scanco Medical AG, Brüttisellen, Switzerland) at a 10-μm resolution. Mineralized tissues were segmented by a global thresholding software. Standardized nomenclature was used for the bone parameters measured. For the femurs, trabecular parameters were measured at the secondary spongiosa of the distal metaphysis and cortical parameters were determined in a 1mm ring at the mid-diaphyseal region according to previously published guidelines⁸⁸. For the 3^rd lumbar vertebra only trabecular parameters were measured. Growth plate thickness was measured in the distal epiphysis of 5-week old mice. Briefly, for each sample, 2D projections from anterior, posterior and mid-longitudinal locations were obtained, the average thickness of the growth plate obtained from the three projections was quantified using the ImagePro Analyzer 7.0 software (Media Cybernetics, Inc.).