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. 2021 Jan 14;10(1):161. doi: 10.3390/foods10010161

Figure 4.

Figure 4

PA-dependent disturbance of ABCC2 driven efflux in HepaRG cells. (A) Differentiated HepaRG cells were incubated for 24 h or 14 days with 35 µM of echimidine, senecionine, or the solvent (1.7% DMSO and 0.35% ACN). To localize the bile canaliculi, the cells were incubated with 5 µM of 5(6)-carboxy-2′,7′-dichloro-fluorescein diacetate (CDFDA) for 30 min at 37 °C, and then analyzed on the fluorescence microscope Axio Observer.D1 (objective EC Plan-Neofluar 5x/0.16 Ph 1) under transmitted light and after excitation with 470 nm at 525 nm. The membrane-bound, non-fluorescent CDFDA is intracellularly converted by esterases into the green fluorescent ABCC2 substrate 5(6)-carboxy-2′,7′-dichlorofluorescein (CDF). By ABCC2-mediated transport, CDF enters the bile ducts. Representative sections are shown. The indicated scale applies to all images. (B) Exemplarily enlarged images of CDF fluorescence after treatment of HepaRG cells for 24 h with the solvent, 35 µM of echimidine, or 35 µM of senecionine. The indicated scale applies to all images of this enlargement.