H&E staining was carried out as the standard protocol described. Paraffin sections were dewaxed, hydrated, and stained with hematoxylin and eosin, respectively, and then dehydrated and mounted. IHC was performed to measure the expression of UCP1 protein in mouse adipose tissues as described previously [47 (link)]. Section was dewaxed and hydrated, followed by antigen retrieval. Endogenous peroxidase was blocked by 3% hydrogen peroxide solution (PV-9000, ZSGB-Bio, Beijing, China). The section was incubated with the blocking goat serum (C0265, Beyotime, Shanghai, China) for 15 min and immunostained with anti-UCP1 antibody (ab10983, Abcam, Cambridge, UK) overnight at 4 °C. After three washes with PBS, the slides were stained with horseradish peroxidase-conjugated anti-rabbit IgG (PV-9000, ZSGB-Bio, Beijing, China) and DAB Chromogenic Kit (ZLI-9018, ZSGB-BIO, Beijing, China), followed by counterstaining with hematoxylin. The full section was scanned by using the Panoramic Scanning Microscope-VS120 (Olympus Life Science, Tokyo, Japan). Cell size and the density of UCP1-positive staining (brownish yellow) was analyzed by Image J.
Free full text: Click here
