Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Organic solute transporter OSTα/β is overexpressed in nonalcoholic steatohepatitis and modulated by drugs associated with liver injury

doi: 10.1152/ajpgi.00310.2017

Figure Lengend Snippet: Expression of organic solute transporter (OST) α/β in human control livers and patients with nonalcoholic steatohepatitis (NASH) and primary biliary cholangitis (PBC). A: quantitative RT-PCR analysis of mRNA levels of OSTα and OSTβ in liver samples of NASH and PBC patients and controls. B and C: Western blots for OSTα and OSTβ proteins and corresponding densitometry. Values are means ± SD. ****P = 0.0001; ***P = 0.0007; **P < 0.005; *P = 0.0218. D: representative immunofluorescence of OSTα (red) and OSTβ (green) proteins in liver samples of NASH and PBC patients. Nuclei of cells in liver samples were stained with 4′,6-diaminido-2-phenylindole (DAPI, blue). Scale bars = 100 μm. Insets: colocalization of OSTα and OSTβ on parenchymal membrane.

Article Snippet: Before immunofluorescence (IF) of liver tissues, the function and specificity of anti-OSTα (Abcam) and anti-OSTβ (Sigma-Aldrich) antibodies were tested with a multitissue human microarray containing kidney cortex, kidney medulla, appendix, lung, colon, pancreas, and liver (Translational Pathology Laboratory, University of North Carolina, Chapel Hill, NC).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Staining

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Organic solute transporter OSTα/β is overexpressed in nonalcoholic steatohepatitis and modulated by drugs associated with liver injury

doi: 10.1152/ajpgi.00310.2017

Figure Lengend Snippet: Expression and function of human OSTα/β in the established OSTab cell line. A: mRNA expression of human OSTα and OSTβ in Flp-In 293 cells transfected with OSTα/β vector (OSTab 293) compared with nontreated Flp-In 293 (wild-type 293) cells, Flp-in 293 cells transfected with empty vector (Mock 293), sandwich-cultured human hepatocytes (SCHH), and SCHH treated with a model bile acid, chenodeoxycholic acid (SCHH-CDCA, 100 μM, 24 h). Expression ratio of OSTα to OSTβ cannot be compared with the given normalization to wild-type. Values are means ± SD. Data were normalized to GAPDH and then compared with wild-type Flp-In 293 cells. B: immunofluorescence showing OSTα (green) and OSTβ (green) protein levels in OSTab and Mock cells. Scale bars = 100 µm. C and D: uptake of taurocholate (TCA) in OSTα/β-overexpressing Flp-In 293 cells (OSTab) and Flp-In 293 cells transfected with empty vector (Mock). C and D: Na+-free buffer and low-pH buffer increased OSTα/β-mediated uptake of TCA. OSTab and Mock cells were preincubated in HBSS-HEPES (pH 7.4) extracellular fluid (ECF) with sodium (ECF Na+, pH 7.4) or ECF in which sodium was substituted with choline (ECF Na−, pH 6.4, 7.4, and 8.4) at 37°C for 10 min. Uptake was initiated by addition of [3H]TCA or 3H-labeled estrone sulfate (300 nCi/ml, 20 µM, 2 min, 37°C) in the corresponding buffer. Values are means ± SD from 3 independent experiments. ****P = 0.0001; ns, not significant.

Article Snippet: Before immunofluorescence (IF) of liver tissues, the function and specificity of anti-OSTα (Abcam) and anti-OSTβ (Sigma-Aldrich) antibodies were tested with a multitissue human microarray containing kidney cortex, kidney medulla, appendix, lung, colon, pancreas, and liver (Translational Pathology Laboratory, University of North Carolina, Chapel Hill, NC).

Techniques: Expressing, Transfection, Plasmid Preparation, Cell Culture, Immunofluorescence, Labeling