Hematoxylin and eosin (HE) and periodic acid Schiff (PAS) staining were performed as follows. The tissues were fixed in 4% paraformaldehyde and subsequently embedded in paraffin. Then, 4-μm-thick cross-sections were processed and stained with HE or PAS for morphological analysis.
Immunohistochemical and immunofluorescent staining were performed as follows. Human or rat tissue was fixed in 4% paraformaldehyde, dehydrated, embedded in paraffin wax, and serially sectioned at a thickness of 4 μm. The sections were incubated with primary antibodies overnight at 4 °C. Subsequently, the sections were stained with fluorescent secondary antibodies. The nuclei were stained with DAPI (Invitrogen). The sections were imaged using a fluorescence microscope (Olympus BX53, Tokyo, Japan). A minimum of 5 random images from 3 samples were analyzed per group. Immunohistochemical statistical analysis was conducted with the Fromowitz comprehensive scoring method [30 (link)]. The details of the antibodies are listed in Additional file1 : Supplementary Table 1.
Immunohistochemical and immunofluorescent staining were performed as follows. Human or rat tissue was fixed in 4% paraformaldehyde, dehydrated, embedded in paraffin wax, and serially sectioned at a thickness of 4 μm. The sections were incubated with primary antibodies overnight at 4 °C. Subsequently, the sections were stained with fluorescent secondary antibodies. The nuclei were stained with DAPI (Invitrogen). The sections were imaged using a fluorescence microscope (Olympus BX53, Tokyo, Japan). A minimum of 5 random images from 3 samples were analyzed per group. Immunohistochemical statistical analysis was conducted with the Fromowitz comprehensive scoring method [30 (link)]. The details of the antibodies are listed in Additional file
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