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. 2023 Dec 9;14:8159. doi: 10.1038/s41467-023-43795-x

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© The Author(s) 2023

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. 5 — A Schematic illustration of organoids transplantation under the kidney capsule of immunodeficient mice either at the RV stage (d14, blue frames) or after differentiation (d24, burgundy frames), created with BioRender.com. Organoids were left to grow in vivo for 14 or 28 days prior to retrieval and analysis. B representative images of kidneys with engrafted AGTR1-/- or Isogenic Control (IC) organoids transplanted at day 24. C IF Stained AGTR1-/- and IC explanted organoids contain PT (HNF4a, LTL), podocytes (NPHS2+) and DT (ECAD, GATA3). D Comparison of the mean area of four explanted AGTR1-/- and four IC d24 organoids. Results are presented as mean ± S.E.M from each line. Comparison was performed using a two-sided t test. n s= not significant; p = 0.6. E Representative images of kidneys with transplanted AGTR1-/- or IC d14 organoids. Only IC organoids engrafted. F Comparison of the mean area of 10/22 explanted AGTR1-/- and 10/22 remnants of IC d14 organoids. Results are presented as mean ± S.E.M from each line. Comparison was performed using a two-sided t-test. ****p < 0.0001. G IF images of engrafted IC d14 organoid. Frame (i) contains tiled 10x images showing a section of the engrafted organoid and mouse kidney. Wire frame is enlarged in (ii, 40x magnification). A different engrafted organoid was stained for TFAP2a in frame (iii). Scale bars in all frames = 100 µm. Q-PCR analyses of VEGF-A gene expression in d14 (H) and day 24 (I) isogenic control (IC), AGTR1-/-, ACE-/- and P-ACE iPSC-derived organoids. Results are presented as mean ± S.E.M of n = 4 biologically independent experiments. Comparisons were performed using one-way ANOVA on ranks. P-values were adjusted for multiple comparisons using the two-stage step-up method of Benjamini, Krieger and Yekutieli. *p < 0.03, ELISA assay to detect VEGF-A protein in conditioned media from d14 (J) and day 24 (K) IC, AGTR1-/-, ACE-/- and P-ACE organoids. Results are presented as the mean ± S.E.M of n = 5 biologically independent experiments. Comparisons were performed using one-way ANOVA on ranks. P-values were adjusted for multiple comparisons using the P-values were adjusted for multiple comparisons using the two-stage step-up method of Benjamini, Krieger and Yekutieli. *p = 0.04, **p = 0.003, **p = 0.001. Abbreviations: ECAD- E-Cadherin (CDH1), GATA3- GATA binding protein 3. Scale bars=100µm. Source data for Fig. 5D, F and H-K are provided as a Source Data file.