Figure 3  Curcumin suppresses lox-1 expression in activated HSCs in vitro
(A). Luciferase activity assays of cells transiently transfected with the lox-1 promoter luciferase reporter pLOX-1-Luc, and treated with curcumin after transfection. Luciferase activities were expressed as relative units after β-galactosidase normalization (means ± s.d.; n≥6). *P<0.05 versus cells without treatment (the first column on the left). The floating schema denoted the luciferase reporter construct pLOX-1-Luc in use and the application of curcumin to the system. (B). Real-time PCR analyses of the steady-state levels of LOX-1 mRNA in cells treated with curcumin. mRNA fold changes were calculated as stated in Materials and Methods. Values were expressed as means ± s.d. (n≥3). *P<0.05 versus the untreated control (the first column on the left); (C). Western blotting analyses of LOX-1 in cells treated with curcumin at indicated concentrations. β-actin was used as an internal control for equal protein loading. Representative was from three independent experiments. Italic numbers beneath the blot were fold changes in the densities of the bands compared to the control without treatment in the blot (n=3), after normalization with the internal invariable control β-actin. (D). Immuno-staining of cultured HSCs treated with or without ox-LDL (10 μg/ml) plus or minus curcumin (10 μM) for evaluating the impact of curcumin on the abundance of intracellular ox-LDL. DAPI in mounting solution was used for staining nuclei. Representative views were presented.