Figure 4

Establishment of in vitro CIDEA reporter primary culture system. Stromal vascular fractions of adipose tissue from homozygous CIDEA reporter mice were isolated, cultured, and differentiated. (a) In vitro bioluminescence imaging of preadipocyte (Pre) and fully differentiated adipocytes (AC) from BAT. Cells were treated with growth medium containing d-luciferin (150 μg/ml). (b) Luciferase assay of cell lysate from (a). (c) Immunofluorescence staining image of differentiated adipocytes from BAT and iWAT. CIDEA (Green) and tdT (Red) double positive cells (CIDEA⁺ tdT⁺) were indicated with a dashed yellow line. Nuclei were counterstained with DAPI (Blue). Bar = 10 μm. (d) Quantification of the number of CIDEA⁺ tdT⁺ cells per field. (e) High magnification images of CIDEA⁺ tdT⁺ adipocytes. Nuclei and lipid were counterstained with DAPI and HCS LipidTOX. Bar = 5 μm. Data were analyzed by an unpaired, two-tailed t test in (b,d) (mean ± SEM; n = 3–4 *P < 0.05, ***P < 0.001).