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

Fig. 4. PT patterning in RAAS deficient kidney organoids is not sensitive to hypoxia.

Fig. 4

A Schematic representation of the protocol used to generate kidney organoid in standard condition (21%O2) and in a hypoxia chamber (2%O2), created with BioRender.com. B Representative confocal images PT markers (LTL, HNF4a, and ASS1) in ACE-/, AGTR1-/-, and P-ACE and their respective Isogenic Control (IC) organoids cultured in standard (21%O2) versus hypoxic (2%O2) conditions. Magnification - 40x; scale bars = 100 µm. C Quantification of PT in ACE-/-, AGTR1-/-, and P-ACE and their respective isogenic control organoids grown either in standard (21%O2) or hypoxic (2%O2) conditions. Bar graphs show the mean ratio of HNF4a (PT cells) to TFAP2b (DT cells) positive cells in mutant organoid compared to controls. Each dot represents the mean of x4 z-sections per organoid. Quantification was performed on n = 28 Isogenic Control (IC), n = 27 ACE-/-, n = 27 AGTR1-/-, and n = 23 P-ACE iPSC-derived organoids grown in 21%O2 and n = 16 IC, n = 25 ACE-/-, n = 23 AGTR1-/- and n = 18 P-ACE iPSC-derived organoids grown in 2%O2 from n = 4 biologically independent differentiation experiments. Data is presented as mean ±S.E.M. Comparisons were performed using a two-sided t test. ns = not significant. p = 0.5 for IC 21%O2 vs 2%O2, p = 0.07 for ACE-/- 21%O2 vs 2%O2, p = 0.7 for AGTR1-/- 21%O2 vs 2%O2 and p = 0.67 for P-ACE 21%O2 vs 2%O2. Abbreviations: LTL- Lotus tetragonolobus lectin, HNF4a- Hepatocyte Nuclear Factor 4α, ASS- ArgininoSuccinate Synthetase. TFAP2b- Transcription factor AP-2 beta. P-ACE- AR-RTD patient-derived iPSC line. Source data for Fig. 4C is provided as a Source Data file.