GEO series

GFP Reporter System Reveals Cell-to-Cell Variability in Aquaporin-2 Expression

GSE310089 Mus musculus (mouse) RNA-seq (gene expression) 22 samples Submitted 2025/11/17 Platform GPL24247

At a glance

RNA-seq (gene expression) · mouse · 22 samples

What it is: RNA-seq (gene expression)Gene activity measured by sequencing. The summary does not mention single-cell work, so this is most likely bulk: one average per sample, the usual starting point for comparing groups.
Organism: Mus musculus (mouse)
Samples: 22 samples: 22 samples, each with a distinct title — no replicated groupsGroups are read from the sample names, not from the paper. Check the list below.
Data you can get: A processed table (series matrix) and supplementary files from NCBI; the raw sequencing reads from ENA. Which of these hold counts depends on the study, so check the file list.
Paper: GFP reporter system reveals cell-to-cell variability in aquaporin-2 expressionAmerican journal of physiology. Cell physiology · 2026

Summary (written by the submitters)

Vasopressin regulates transcription of the aquaporin-2 gene (Aqp2) in collecting duct principal cells. To investigate regulatory mechanisms in Aqp2 gene transcription, we engineered an Aqp2 reporter cell line using CRISPR/Cas9 to insert a green fluorescent protein (GFP) cassette at the endogenous Aqp2 gene locus in mpkCCD cells. In the absence of dDAVP, a vasopressin analog, these cells exhibited low or undetectable GFP and Aqp2 expression in all cells.

Read the full summary ↓

dDAVP stimulation (1nM dDAVP for 48hrs) markedly increased both GFP and Aqp2 expression together with reversal upon dDAVP removal. These observations demonstrate that GFP faithfully tracks Aqp2 expression. Interestingly, fewer than 50% of cells express GFP and Aqp2 after dDAVP or forskolin, indicating significant variability even though they were clonally-derived. We flow-sorted the GFP- cells (Aqp2-) and GFP+ cells (Aqp2+), regrew them, and restimulated them separately with dDAVP. Cells originating from GFP- cells gave rise to both GFP- cells and GFP+ cells, and GFP+ cells similarly regenerated both GFP- and GFP+ populations in the same proportion. Flow cytometry analysis of the DNA content showed variability in cell cycle phases, with most GFP+ cells in G0/G1, and more GFP- cells in G2/S. RNA-seq analysis of the GFP- and GFP+ cells revealed increased abundance of cell-cycle related transcripts in the GFP- cells. We conclude that: 1) heterogeneity in Aqp2 expression is related to cell cycle state; and 2) the newly generated reporter cell line will likely serve as a useful tool to study Aqp2 transcriptional regulation.

Published in

GFP reporter system reveals cell-to-cell variability in aquaporin-2 expression

Chen L, Murillo-de-Ozores AR, Park E et al. · American journal of physiology. Cell physiology 2026 · PMID 41730290 · doi:10.1152/ajpcell.00936.2025

This dataset

Open GSE310089 in finder → Find more mouse RNA-seq datasets →

Download · which file do I need?

Direct links to NCBI and ENA, no account and no request form. Pick by what you want to do: take the small processed table to look, the supplementary files to re-analyse, or the original raw data to start over.

1 · Just have a look

Processed table

The series matrix: one table of values per sample as the submitters processed it. The smallest download, and it opens in Excel or R. Start here to see what the data look like.

Series matrix ↓

2 · Re-analyse it

Supplementary files

The count tables and other files the authors uploaded. Take only the samples you need, or the whole study as GSE310089_RAW.tar (it can run to tens of gigabytes).

All supplementary files ↗ Whole study · RAW.tar ↓

3 · Start from the raw reads

Raw sequencing reads

The original FASTQ files, straight from the sequencer, held at ENA. Large, and you align them yourself, so only choose this if you need to re-process the study from scratch.

Raw reads on ENA ↗

What the files usually look like: For RNA-seq the counts are usually a table among the supplementary files (look for 'counts', 'expression', 'FPKM' or 'TPM' in the file name). The series matrix of a sequencing study is sometimes nearly empty.

Also filed as BioProject PRJNA1365104 and SRA study SRP645362. Searching any of these in the dataset finder brings you back here.

NCBI GEO page ↗ Paper (PMID 41730290) ↗

Study design

22 samples, each with a distinct title — no replicated groups

Any grouping that defines a comparison lives in the sample characteristics, which GEO records separately from the titles.

How we read this: Read from 22 sample titles: 22 distinct titles with little repetition. These groups are guessed from the sample names, not taken from the paper, so check them against the sample list below before relying on them.

Samples in this study

GSM9287949 Replicate 1, WT-mpkCCD cell, no dDAVP
GSM9287950 Replicate 2, WT-mpkCCD cell, no dDAVP
GSM9287951 Replicate 3, WT-mpkCCD cell, no dDAVP
GSM9287952 Replicate 1, WT-mpkCCD cell, dDAVP
GSM9287953 Replicate 2, WT-mpkCCD cell, dDAVP
GSM9287954 Replicate 3, WT-mpkCCD cell, dDAVP
GSM9287955 Replicate 1, GFP reporter C1 cell, GFP negative, dDAVP
GSM9287956 Replicate 2, GFP reporter C1 cell, GFP negative, dDAVP
GSM9287957 Replicate 1, GFP reporter C1 cell, GFP positive, dDAVP
GSM9287958 Replicate 2, GFP reporter C1 cell, GFP positive, dDAVP
GSM9287959 Replicate 1, GFP reporter C11 cell, GFP negative, dDAVP
GSM9287960 Replicate 2, GFP reporter C11 cell, GFP negative, dDAVP
GSM9287961 Replicate 1, GFP reporter C11 cell, GFP positive, dDAVP
GSM9287962 Replicate 2, GFP reporter C11 cell, GFP positive, dDAVP
GSM9287963 Replicate 1, GFP reporter C43 cell, GFP negative, dDAVP
GSM9287964 Replicate 2, GFP reporter C43 cell, GFP negative, dDAVP
GSM9287965 Replicate 1, GFP reporter C43 cell, GFP positive, dDAVP
GSM9287966 Replicate 2, GFP reporter C43 cell, GFP positive, dDAVP
GSM9287967 Replicate 1, GFP reporter C50 cell, GFP negative, dDAVP
GSM9287968 Replicate 2, GFP reporter C50 cell, GFP negative, dDAVP
GSM9287969 Replicate 1, GFP reporter C50 cell, GFP positive, dDAVP
GSM9287970 Replicate 2, GFP reporter C50 cell, GFP positive, dDAVP

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