Mouse Femur Histological Analysis

The mouse femurs were harvested and fixed in 4% paraformaldehyde for 2 days, and decalcified uisng 10% ethylenediaminetetraacetic acid, embedded in paraffin, then cross-sectioned (5 μm) on the rotary microtome (RM2235, Leica, Germany). Hematoxylin and eosin (HE) staining was conducted as the previous study (Zhang et al. 2018 (link)). The differentiation of osteoclasts was measured by TRAP staining. Bone sections were labeled with Masson staining according to the manufacturer’s protocol (Sigma-Aldrich, Missouri, USA). Immunohistochemical (IHC) staining was performed as previously described (Zhao et al. 2021 (link)). The primary antibody against NFATc1 (sc-7294, 1:200) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Five fields were randomly selected per section and photographed.

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Yu X., Rong P.Z., Song M.S., Shi Z.W., Feng G., Chen X.J., Shi L., Wang C.H, & Pang Q.J. (2021). lncRNA SNHG1 induced by SP1 regulates bone remodeling and angiogenesis via sponging miR-181c-5p and modulating SFRP1/Wnt signaling pathway. Molecular Medicine, 27, 141.

Publication 2021

RM2235 Masson staining sc-7294

Antibody Bone Embedded paraffin Eosin Ethylenediaminetetraacetic acid Femurs Hematoxylin Microtome Mouse Osteoclasts Paraformaldehyde

Corresponding Organization :

Other organizations : University of Chinese Academy of Sciences, Ningbo University

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Variable analysis

independent variables

Treatments (e.g., drug, intervention) applied to the mice

dependent variables

Differentiation of osteoclasts (measured by TRAP staining)
Bone structure (assessed by Masson staining)
Expression of NFATc1 (measured by immunohistochemical staining)

control variables

Mouse femurs
Fixation in 4% paraformaldehyde for 2 days
Decalcification using 10% ethylenediaminetetraacetic acid
Embedding in paraffin
Cross-sectioning (5 μm) on the rotary microtome
Hematoxylin and eosin (HE) staining

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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