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Fig. 7. Hep G2 cells containing apical Na+/K+-ATPase are defective in canalicular excretion. (A) Polarized Hep G2 cells were treated with 20 µg/ml tunicamycin for 24 hours (TM) or untreated (CTL). Fluorescein diacetate (FDA, green) was added to the culture media for 30 minutes; the locations of bile canaliculi were visualized by F-actin staining (red). The bile canaliculi capable of excreting FDA via transcytotic transport were stained yellow (arrows), or stained red otherwise (arrowheads). (B) Fluorescently labelled 3 kDa dextrans (green) were added to the culture media for 1 hour; the locations of bile canaliculi were visualized by F-actin staining (red). The fluorescent dextran could enter the canalicular lumen via permeable tight junctions between the neighbouring cells (the paracellular pathway). Bile canaliculi active in paracellular transport were stained yellow (arrows), and stained red if inactive (arrowheads). (C) Quantification of the bile canaliculi (BC) apical domains active in transcytotic (grey bars) or paracellular transport (open bars) in the presence (TM) or absence (-) of tunicamycin treatments. More than 500 apical domains were counted for each experiment and results show the means ± s.d. *P<0.05 by Student's t-test. Bars, 20 µm.
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