Fig. 7

FACI promotes cellular LDL uptake. A Images of TIRFM (left) and spinning disc microscopy (right) showing the colocalization of mRuby2-FACI and EGFP-LDLR to PM and cytosol, respectively. Scale bar, 10 μm. B Fluorescent LDL uptake assay in WT and FACI^−/− HeLa cell lines. The amounts of endocytosed pHrodo-Red-LDL were measured by flow cytometry. A representative flow cytometric histogram shows pHrodo-Red-LDL uptake in WT and FACI^−/− HeLa cell lines (left). Quantitative analysis of pHrodo-Red-LDL uptake was performed in replicated experiments (right). Data are mean fluorescence intensity ± SD. **: P < 0.01 by two-tailed paired Student’s t-test. C Fluorescent LDL uptake was compared between FACI-expressed (Dox group) and control (no Dox group) AML12 cells. The amounts of endocytosed pHrodo-Red-LDL were measured by flow cytometry. The representative flow cytometric histogram (left) and quantitative analysis (right) were indicated. Data are mean fluorescence intensity ± SD. *: P < 0.05 by two-tailed paired Student’s t-test. D-G C57BL/6 J mice (8 weeks old, n = 6) and FACI-/- mice (8 weeks old, n = 7) were fed with a high-cholesterol diet for 4 weeks. Plasma total cholesterol (TC; D), LDL-cholesterol (LDL-C; E), HDL-cholesterol (HDL-C; F) and triglyceride (TG; G) were compared. *: P < 0.05 by two-tailed paired Student’s t-test. H-I C57BL/6 J mice (6 weeks old) were injected with AAV-FACI (n = 8) or AAV-GFP (n = 7). AAV-FACI is an adeno-associated recombinant virus (AAV) expressing FACI driven by a liver-specific promoter and enhancers. After two-week transduction, mice were fed with a high-cholesterol diet for another 4 weeks. Mice were sacrificed and liver total mRNA was extracted. FACI expression in livers was analyzed by RT-qPCR (H) and immunoblotting (I). Plasma total cholesterol (TC; J), LDL-cholesterol (LDL-C; K), HDL-cholesterol (HDL-C; L) and triglyceride (TG; M) were compared between AAV-FACI and AAV-GFP mice. *: P < 0.05. ***: P < 0.001 by two-tailed paired Student’s t-test