DE cells were plated and differentiated in 24-well-plates and were stained with BODIPY 493/503 and DAPI to assess lipid accumulation/content. Cells were fixated in 4% paraformaldehyde for 15 min at room temperature, washed twice in PBS and then stained with 10 μg/ml Bodipy 493/503 (lipid droplets) and 1 μg/ml DAPI (nuclear staining) for 15 min. Lastly, cells were washed and imaged with Zeiss Cell Observer microscope with a 10X objective. 9 randomly selected fields of view per well were imaged and all images were acquired with exact same settings. Analysis of the bodipy area and intensity and cells surface area was performed in ZEN software. The relative area/intensity was calculated by dividing the Bodipy area or intensity for the area occupied by the cells (cells surface area) in each field of view.
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