Fig 1: GPR182 mediates lipoprotein transport.(A and B) GPR182+ CHO and control WT CHO cells were incubated with Dio-HDL (A) or Dil-CM (B) at 37°C to assess lipoprotein endocytosis. GPR182 mAb (clone 1A5) was added to assess its blocking capacity. Cells were costained with WGA before imaging. (C) Dil-HDL uptake was assessed in WT HEK293T cells, SR-B1-KO cells, and SR-B1–KO cells transfected with GPR182 at 37°C. (D) Dil-LDL uptake was assessed in WT HEK293T cells, LDLR-KO cells, and LDLR-KO cells transfected with GPR182 at 37°C. (E and F) SVEC4-10 cells were treated with control or GPR182 mAb before incubation with Dio-HDL (E) or Dil-CM (F) at 37°C for 30 minutes. Cells were then costained with WGA to assess lipoprotein endocytosis. (G) SVEC4-10 cells, with or without PCSK9 treatment, were incubated with control or GPR182 mAb before culturing with Dil-LDL at 37°C for 30 minutes. Cells were then costained with WGA to assess lipoprotein endocytosis. (H) Dil-labeled HDL or CM was added into the inserts at 37°C. After overnight incubation, supernatant from the lower chambers was collected, and fluorescence was quantified using a fluorescence plate reader (n = 3 or 7). All experiments were repeated 2–3 times. Data represent the mean ± SEM. One-way ANOVA was used to compare the means of more than 2 groups (A and B). An unpaired, 2-tailed Student’s t test was used to compare the means of 2 groups (E–H). *P < 0.05 and ****P < 0.0001. Scale bars: 10 μm.