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. 2024 Oct 22;10:106. doi: 10.1038/s41421-024-00708-6

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© The Author(s) 2024

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 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. 5 — a Illustration of the establishment of orthologous NUMB^R632H mutation knock-in mice. b Representative electropherograms show the generation of mice heterozygous (WT/RH) or homozygous (RH/RH) for the introduced mutation (blue shadow). c Chi-square to compare observed and expected distributions of three genotype according to Mendel’s law. d–g Comparison of the ratio of urinary uric acid to urinary creatinine (d, WT/WT n = 8; RH/RH n = 10; t = 2.600, df = 16), the fractional excretion of uric acid (FEUA) (e, WT/WT n = 5; RH/RH n = 8; t = 2.570, df = 11), serum urate (f, WT/WT n = 6; RH/RH n = 8; t = 2.272, df = 12) and creatinine clearance (g, WT/WT n = 8; RH/RH n = 8; t = 0.5308, df = 14) between in WT/WT mice and in NUMB^R632H/NUMB^R632H (RH/RH) mice. h Representative images of immunofluorescence staining of NUMB in kidney tissue of WT/WT mice and NUMB^R632H /NUMB^R632H mice. The solid lines indicate the edge of renal tubules, and the dashed lines mark the lumen of renal tubules. T: renal tubules; Ap: apical side of renal tubular epithelia cells (RTECs); Ba: basolateral side of RTECs. White arrows indicate sporadic distribution of granule-like structure of NUMB^R632H protein in cytoplasm of RTECs. i The statistic comparison of NUMB protein (top panel), apical NUMB (middle panel) and NUMB aggregate-like structure (bottom panel) each renal tubule between in WT/WT mice (n = 7) and in NUMB^R632H/NUMB^R632H mice (n = 6). j Representative images of immunofluorescence staining of ABCG2 and basolateral marker Na/KATPase. The white arrow heads indicate the distribution of ABCG2 (red). In WT/WT mice, ABCG2 is distributed to the apical side just beneath the LTL signal. However, in NUMB^R632H /NUMB^R632H mice, ABCG2 located to the basolateral side with increased co-localization with Na/K ATPase. The illustration of ABCG2 distribution is shown in the bottom left, and the statistic result of the percentage of apical ABCG2 is shown in the bottom right (n = 6 per group, t = 19.34, df = 10). Statistics was calculated using two-tailed unpaired Student’s t test. The results are presented as mean ± SD.