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Human tissue and mouse bone sections were used as positive and negative controls, respectively, for human-specific antibodies.
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Sections of mouse femur were used as negative controls to rule out any cross-reactivity of the antibodies with mouse tissue, and human bone sections served as positive controls (supplementary material Fig. S3).
We chose to further analyze Cox-2 by immunohistochemical (IHC) analysis using Cox-2 antibody and mouse embryonic (E17.5) long bone sections.
To corroborate this finding, we performed in situ hybridization on long bone sections from mouse embryos of later stages.
This implies that understanding the precise pathomechanisms underlying the skeletal defects in this large group of inherited disorder is of importance, and that a profound histomorphometric analysis should be performed on bone sections from mouse models and/or patients with defined lysosomal storage disorders.
We also established the modified protocols for in situ hybridization and BrdU labeling of bone sections from mice.
In addition, we established the modified methods for in situ hybridization and BrdU labeling of bone sections from mice to study osteoclast survival in vivo.
Furthermore, we established the optimal protocols for in situ hybridization and BrdU labeling of bone sections from mice to study osteoclast life span in vivo.
Unstained, electron-dense granules in 6-day-old mouse rib have been observed in bone sections prepared under non-aqueous conditions [48].
Micro-computed tomography of PC5/6-knockout embryos at E18.5 confirmed a reduction in mineralized bone, and in situ hybridization performed on cryo-sections of normal mouse bone using Pcsk5 and Opn anti-sense and control-sense cRNA probes indicated the co-localization of the expression of these genes in bone cells.
This is in sharp contrast to the observations of Aqeilan et al [16] who observed consistently higher numbers of osteoclast activity in bone sections of KO mice however they did not perform quantitative analyses in those studies.
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