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. 2020 Jun 2;11:2767. doi: 10.1038/s41467-020-16525-w

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 7 — a Western blots of jejunum homogenate from WT (n = 8) and Q140K+/+ (n = 6) mice showed a significant 78% decrease in abundance of the Q140K+/+ protein (p = 0.0046; loading constant determined by total lane protein analysis; ±SEM). b Quantitative real-time PCR analysis of small intestine mRNA shows no significant difference in the mRNA of ABCG2 in WT as compared to Q140K+/+ (n = 5 for both, p = 0.5132; ±SEM). c Representative immunofluorescence micrographs of fixed sections of mouse small intestines (from n = 3 WT and n = 3 Q140K+/+ mice) stained for ABCG2 (green), villi brush border marker beta-actin (red), and colocalization of ABCG2 and beta-actin (yellow) at 200×(all scale bars 10 μM); quantification of immunofluorescent ABCG2 signal in either WT and Q140K+/+ jejunum show significant differences in [PTP/TP] (p = 0.018; n = 4 analysis areas from jejunum sections from 4 WT animals and n = 3 analysis areas from 3 Q140K+/+ animals; ±SEM). d Image of intestinal loops prepared from mouse small intestines; and graphic demonstrating the functional lumen urate accumulation assay. e Plot of urate concentration versus luminal UA flux fit successfully with a Michaelis–Menten curve, Vmax = 59.58 ± 2.74 μM/cm²/min; Km = 48.45 ± 11.44μM, n = 5 animals for each of seven urate concentrations; ±SEM. f Mean luminal flux in WT (n = 17) and Q140K+/+ (n = 10) jejunum loops were significantly different (p < 0.0001); and in colonic loops (n = 6 for both; p = 0.974). g Modeled ABCG2 mediated flux (mean ABCG2 inhibitor TPX sensitive flux subtracted, see “Methods” section) demonstrate almost complete loss in Q140K+/+ animals (reduced 84.2%; WT n = 17 and Q140K+/+ n = 10, p < 0.0001; ±SEM). h Calculated jejunum urate net transport (see “Methods” section) shifts from net luminal secretion to net absorption in the Q140K+/+ loops (WT n = 5; Q140K+/+ n = 3; p = 0.0036; ±SEM). Statistical analysis: a–c, f–h, two-tailed Student’s t-test. Source data are provided as a Source data file.