Fig. 7. Blocking endocytosis inhibits lipid droplet growth and HFD-induced obesity.

a On the day of experiment, WT and Cd36^−/− adipocytes were pretreated with PP2 (20 μM), ML348 (10 μM), or piceatannol (40 μM) for 1 h, then treated with oleate (100 μM) and BODIPY 493/503 (0.1 μg/ml), and subjected to live imaging on a Zeiss LSM-780 confocal microscope in a 3D Z-stack mode for 2 h. The number of cells was 12, 9, 8, and 10 for DMSO, PP2, ML348, and piceatannol-treated WT cells, and 8, 8, 8, and 10 for corresponding groups in Cd36^−/− cells, respectively. Total volume of lipid droplets in each cell was calculated. Each value represents mean ± SEM, and the volume at 0 min was normalized to 1.0. Two-sided Student’s t test was performed between DMSO- and PP2-, ML348- or piceatannol-treated cells. b–d WT and Cd36^−/− mice (8-week-old male, n = 6/group) were daily gavaged with vehicle (0.5% methyl cellulose), bafetinib (20 mg/kg), or entospletinib (10 mg/kg) at 7 pm and subjected to HFD feeding for 8 weeks. b Body weight of the mice was monitored every week. c, d Gonadal WAT was subjected to H&E staining, and representative pictures were shown (c). Scale bar, 100 μm. Quantification of the surface area of adipocytes in gWAT by ImageJ. The size of each adipocyte was quantified by ImageJ and plotted as mean ± SEM from 287, 488, 305, 324, 362, and 218 cells, respectively d. Two-sided Student’s t test was performed between vehicle and bafetinib (Baf.), or entospletinib (Ent.) treated group, respectively. These experiments were repeated twice. Source data are provided as a Source Data file.