Oil-Red-O (ORO, O0625, Sigma-Aldrich, USA) was used to measure the amount of lipid droplets in a tissue, as we previously reported [11 (link)]. Samples were immersed in ORO (2 mM in 100% isopropanol, 10 µl/mg) and incubated at room temperature for 2 h. ORO-stained tissues were washed in 1% SDS overnight and lipids were extracted by soaking in 100% isopropanol for 4 h, then the extracted fluorescence was measured with a Micro plate reader (SpectraMax Plus384, Molecular Devices, Sunnyvale, CA, USA) at 500 nm.
DiI (468495, Sigma-Aldrich, USA) was used to measure the amount of phospholipids in a tissue. The tissue was immersed in 100% dimethyl sulfoxide (DMSO) for 24 h to allow homogenous loading of DiI deep into the tissue, after which it was again immersed in 0.2 mM DiI in DMSO overnight. Then, the tissue was washed with 1% SDS overnight, followed by the extraction of lipids by soaking in 100% isopropanol for 4 h. The extracted fluorescence was measured using a Micro plate reader (SpectraMax Plus384, Molecular Devices, Sunnyvale, CA, USA) at 550 nm.
DiI (468495, Sigma-Aldrich, USA) was used to measure the amount of phospholipids in a tissue. The tissue was immersed in 100% dimethyl sulfoxide (DMSO) for 24 h to allow homogenous loading of DiI deep into the tissue, after which it was again immersed in 0.2 mM DiI in DMSO overnight. Then, the tissue was washed with 1% SDS overnight, followed by the extraction of lipids by soaking in 100% isopropanol for 4 h. The extracted fluorescence was measured using a Micro plate reader (SpectraMax Plus384, Molecular Devices, Sunnyvale, CA, USA) at 550 nm.
