The diameter and number of adipocytes were determined in 3-4 paraffin-embedded sections of the SAT. The sections were stained with H and E. After staining, the diameters of adipocytes were determined in three sections from young (n = 100 adipocytes) and old rats (n = 100 adipocytes), using the NDP. VIEW image software of NanoZoomer S60(HAMAMATSU, Japan). The number of adipocytes in the same area was counted from three sections from the two groups of rats (n = 3) using the NDP. VIEW image software.
The TUNEL assay was performed in each group (n = 3) using the Apoptotic Detection kit (Cat# MK500, Takara, Japan). All stained sections were imaged under a confocal microscope (Nikon, Japan).
The immunofluorescence assay was performed as previously described (Jeong et al., 2020 (link)). Briefly, sections (n = 3) were incubated in primary antibodies Cleaved Caspase-3 (Cat#9664, CST, United States) at 4 °C overnight and with a secondary antibody for 1 h at room temperature. All stained sections were imaged under a confocal microscope (Nikon, Japan).
The TUNEL assay was performed in each group (n = 3) using the Apoptotic Detection kit (Cat# MK500, Takara, Japan). All stained sections were imaged under a confocal microscope (Nikon, Japan).
The immunofluorescence assay was performed as previously described (Jeong et al., 2020 (link)). Briefly, sections (n = 3) were incubated in primary antibodies Cleaved Caspase-3 (Cat#9664, CST, United States) at 4 °C overnight and with a secondary antibody for 1 h at room temperature. All stained sections were imaged under a confocal microscope (Nikon, Japan).
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