Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2023 Sep 20;621(7980):857–867. doi: 10.1038/s41586-023-06549-9

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 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 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/.

PMC Copyright notice

Fig. 4 — a, UMAP of 449 ionocytes coloured by subcluster. b, Type A, B and C ionocyte gene expression signatures showing relative expression (Z-score of log₂(TPM + 1)). c, Distribution of expression levels (log₂(TPM + 1)) for ionocyte subtype markers (white circle, mean; error bars, 95% confidence interval; n = 449 ionocytes from 8 donors; P value, Wilcoxon). d, Ingenuity pathway analysis of differentially expressed ionocyte subtype genes showing top significantly associated diseases and functional pathways (right-tailed Fisher’s exact test). e, RNAscope validation of ionocyte subtypes using cytospun samples from ALI cultures. Representative image of n = 36 ionocytes. f, Differentiated FOXI1-Cre^ERT2::ROSA-TG ALI cultures induced with OH-Tam and later pulsed-labelled with EdU. Cultures were then stained for Ki67 or EdU. Representative image of n = 20 ionocytes. g, Osmosensory ion and water channel gene expression in different cell types. h, Hyperosmotic and hypoosmotic conditions induce opposing forces on ASL hydration. Left, differentiated ALI cultures were exposed to hyperosmotic (+77 mOsm) and hypoosmotic (−77 mOsm) basolateral media for 24 h and ASL height was measured at 0 and 24 h following small volume addition to the apical surface. Data are mean ± s.e.m. Statistical significance by paired two-tailed Student’s t-test (n = 4 donors, each using 2 cultures). Right, quantification of ionocyte numbers in 21 days ALI cultures maintained under hyperosmotic and hypoosmotic conditions throughout basal cell differentiation. Data are mean ± s.e.m. Statistical significance by paired two-tailed Student’s t-test (hyperosmotic: n = 5 donors, 1 culture per donor, quantified as FOXI1-Cre^ERT2 EGFP⁺ cells; hypoosmotic: n = 4 donors, each using 2–3 cultures, quantified as ATP6V1G3⁺ cells). i, Changes in subtype marker gene expression by RT–qPCR in ALI cultures maintained under hypoosmotic or hyperosmotic stress as in h. mRNA fold change was calculated by the ΔΔC_t method. Statistical significance was determined by paired two-tailed Student’s t-test (n = 4 donors, each using 3 cultures). TPM, transcripts per million.

Source Data