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. 2021 Jul 23;7(30):eabg1371. doi: 10.1126/sciadv.abg1371

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Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).

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Fig. 6 — (A) Differential expression of genes related to bile metabolism and lysosomal functions in fxr^−/− cells as compared to fxr^+/+ cells in cluster 17. (B) qRT-PCR comparing expression of LRE markers in dissected intestines of gender- and size-matched adult fxr^+/+ or fxr^−/− zebrafish. The results are represented as relative expression levels normalized to the housekeeping gene ef1a (mean ± SEM). Statistical significance was calculated by unpaired t test. Representative data from two independent experiments are shown. (C) Confocal single-plane image of the intestinal epithelium in 7-dpf live larvae expressing the Tg(-1.7fabp6:GFP) transgene (green) following gavage with Alexa Fluor 647 dextran (magenta). White arrowheads mark GFP-expressing cells that uptake dextran. The schematic diagram depicting gavage approach for labeling lysosome-rich enterocytes (LREs) in the larval zebrafish intestine using fluorescent luminal cargo is shown in the top panel. Image is representative of six larvae examined. (D) Confocal three-dimensional projections of the intestinal epithelium of 7-dpf larvae expressing the Tg(-1.7fabp6:GFP) (green) and TgBAC(lamp2:RFP) (red) transgenes. White arrowheads mark cells showing both GFP and RFP (red fluorescent protein) expression. Image is representative of five larvae examined. (E) Confocal single-plane image of the intestinal epithelium in 7-dpf fxr^+/+ (top) and fxr^−/− (bottom) larvae expressing the Tg(-4.5fabp2:DsRed) (red) and Tg(-1.7fabp6:GFP) (green) transgenes following gavage with Alexa Fluor 647 dextran (magenta). White arrowheads mark the ileocyte region, which persists in fxr^−/− larvae despite loss of GFP expression. Images are representative of five fxr^+/+ and five fxr^−/− larvae examined. Asteroids mark zebrafish lumen in (C) to (E). Scale bars, 25 μm. (F) Uptake profiles along LRE region following gavage with Alexa Fluor 647 dextran (1.25 mg/ml) in 7-dpf fxr^+/− (n = 14) and fxr^−/− (n = 22) larvae. (G) DsRed fluorescence along the intestine of 7-dpf fxr^+/− (n = 18) and fxr^−/− (n = 24) Tg(-4.5fabp2:DsRed) larvae.