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. 2020 Sep 21;11:4765. doi: 10.1038/s41467-020-18565-8

Fig. 1. FAs trigger internalization of CD36.

Fig. 1

a, b On day 8 of differentiation, 3T3-L1 adipocytes were pretreated with serum-free medium for 4 h, and then treated with BSA-conjugated oleate (100 μM) for indicated time. a One set of cells was subjected to immunostaining with anti-CD36 and anti-CAV1 antibodies. LipidTOX was used to label lipid droplets. Images were taken under a Zeiss LSM-780 microscope in a 3D Z-stack mode and reconstructed using Imaris 9.2.0. b The other set of cells was subjected to surface biotinylation assay and blotted with indicated antibodies. c, d On day 4 of differentiation, 3T3-L1 cells were infected with lentivirus encoding scrambled shRNA or shRNAs against CD36 or CAV1. On day 5, cells were selected with 5 μg/ml puromycin. On day 8, cells were pretreated as in (a) and treated with oleate (100 μM) for 4 h, followed by immunostaining with anti-CD36 and anti-CAV1 antibodies (c), or surface biotinylation assay (d). e, f 3T3-L1 adipocytes were pretreated as in (a) and treated with BSA-conjugated FAs with different chain lengths or saturation (100 μM) for 4 h. Cells were subjected to immunostaining with anti-CD36 antibody (e), or surface biotinylation assay (f). After oleate treatment for 4 h, 3T3-L1 adipocytes were switched to serum-free medium for indicated time and harvested for immunostaining (g) and surface biotinylation (h). The scale bars were as indicated in each figure. These experiments were repeated at least three times. Source data are provided as a Source Data file.