Figure 6
From: Functional polarization of human hepatoma HepaRG cells in response to forskolin
Effects of PXR silencing on FSK-induced formation of BC. (a,b) Untransfected (UNT) and SiNT- or SiPXR-transfected HepaRG cells as well as PXR-KO and control F5 HepaRG clone cells were either exposed to 0.1% (vol/vol) DMSO (control/CTR) or treated by 50 µM FSK for 72 h. (a) Cells were then observed by phase contrast microscopy, allowing to distinguish refractive bright/white BC, indicated by white arrows on phase-contrast microscopic pictures. (b) BC numbers were next quantified by image analysis, as indicated in Methods; data shown are the means ± SEM of at least three independent assays. *p < 0.05 when compared to control cells not exposed to FSK (c,d) HepaRG PXR-KO cells were cultured for 18 days. (c) Cells were then observed by phase contrast microscopy, allowing to distinguish refractive bright/white BC, indicated by white arrows on phase-contrast microscopic pictures. (d) The biliary markers P-gp and MRP2 were immunolocalized, as described in Methods; specific immunolabelling appears as green fluorescence on fluorescence microscopy pictures, whereas DAPI-stained nuclei are blue; BC-associated green fluorescence is indicated by white arrows; the data shown are representative of three independent assays. (a,c,d) White bar = 50 µm.
