Fig. 3. Vectorial hepatic cargo secretion from polarized HLCs.
a Heatmap of Z score-normalized CPM values for enzyme and transporter genes involved in lipoprotein metabolism in nonpol- compared with pol-HLCs (n = biological replicates). b Density distribution of ApoB100 and ApoE secreted from nonpol- and pol-HLCs, labeled metabolically for 4 h with [35S] methionine/cysteine. Supernatants from labeled cells were subjected to density gradient centrifugation followed by ApoB100 or ApoE immunoprecipitation of each fraction, separation by SDS-PAGE, and detection by [35S] fluorography. Results are representative of three independent differentiations. c Nonpol- and pol-HLCs were treated with indicated concentrations of the MTP inhibitor Lomitapide, prior to metabolic labeling, ApoB100-immunoprecipitation and detection. Results are representative of three independent differentiations. d Heatmap for Z score-normalized CPM values of enzyme and transporter genes involved in bile acid metabolism in nonpol- (Non) compared with pol-HLCs (Pol) (n = biological replicates). e Total bile acid release from nonpol- and pol-HLCs treated for 24 h with 10 μg/ml high-density lipoprotein (HDL) or 10 μm cyclosporine A (CsA), as indicated (n = biological replicates). f–g LC-MS analysis of primary and conjugated cholic acid (f) and chenodeoxcycholic acid (g) released from pol-HLCs treated with DMSO or 10 μm 7α-hydroxylcholesterol (7α-CHO) for 4 h (n = biological replicates). Statistical analysis was performed using a two-tailed unpaired t test with Bonferroni adjustment for multiple comparisons.