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. 2023 Feb 7;475(4):489–504. doi: 10.1007/s00424-023-02792-1

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© The Author(s) 2023

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — Identification and characterization of human SVCT1 as a urate transporter. All uptake assays were conducted using transiently SVCT1-expressing HEK293 cells 48 h after plasmid transfection in Krebs–Ringer buffer. Experimental conditions: pH 7.4 unless otherwise indicated, or pH 10 (h); [1-¹⁴C]-vitamin C in the transport buffer was 20 μM (c, j); incubation time and [8-¹⁴C]-urate in the transport buffer were 5 min and 10 μM, respectively, unless otherwise indicated. a–c Functional expression of SVCT1 in HEK293 cells 48 h after the transfection. (a) Immunoblot detection of SVCT1 protein in whole-cell lysates. (b) Intracellular localization of SVCT1 detected by confocal microscopy. Magnified images of representative cells denoted by yellow squares are demonstrated. Scale bars: 10 μm. (c) [1-¹⁴C]-Vitamin C transport activities into the cells. d–i SVCT1 as a urate transporter. (d) Sodium-dependency in SVCT1-mediated urate transport. (e) Insignificant effect of an alkaline pH condition (pH 10) on the urate transport activity of SVCT1. (f) Time-dependent [8-¹⁴C]-urate incorporation into SVCT1-expressing or mock (control) cells. (g) Time profile for SVCT1-mediated [8-¹⁴C]-urate uptake into HEK293 cells, which was calculated by subtracting the urate transport activity of mock cells from that of SVCT1-expressing cells. (h) Concentration dependence in SVCT1-mediated [8-¹⁴C]-urate transport. Regarding the estimated Michaelis-Menten constant (K_m) and maximal velocity (V_max), the 95% confidence interval values were provided in parentheses. (I) Concentration-dependent inhibition of SVCT1-mediated urate transport by vitamin C. IC₅₀, the half-maximal inhibitory concentration. j Inhibitory effect of urate on SVCT1-mediated vitamin C transport at physiological concentrations. Values are shown as % of vehicle control (i, j). Data are expressed as the mean ± SD; where vertical bars are not displayed, the SD was contained within the limits of the symbol; n = 3. ^††P < 0.01 (two-sided t-test; c, e); ns, not significantly different between groups; ^**P < 0.01 vs. the other groups (Tukey–Kramer multiple-comparison test; d); ^##P < 0.01 vs. vehicle control (Williams’ multiple-comparison test; j)