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SRX23880747: GSM8136445: Tcf21-MCM/fl baseline; scRNA-seq; Mus musculus; RNA-Seq
4 ILLUMINA (Illumina NovaSeq 6000) runs: 608.8M spots, 84G bases, 24.5Gb downloads

External Id: GSM8136445_r1
Submitted by: Cincinnati Children's Hospital Medical Center
Study: Transcription factor 21 regulates cardiac myofibroblast formation and fibrosis [scRNA-Seq]
show Abstracthide Abstract
Transcription factor 21 (TCF21) is a basic helix-loop-helix protein required for developmental specification of cardiac fibroblasts from epicardial progenitor cells that normally surround and invade the heart. In the adult heart, TCF21 is expressed in tissue-resident fibroblasts but is downregulated in response to injury or stimuli leading to myofibroblast differentiation. These findings led to the hypothesis that Tcf21 could be a regulator of fibroblast cell fate in the adult mammalian heart and contribute to cardiac fibrosis. Here, single-cell RNA-sequencing was used to study the effect of loss of Tcf21 in cardiac fibroblasts in adult mouse hearts at baseline and after an ischemic cardiac injury (myocardial infarction). Overall design: To investigate the function of TCF21 in vivo, TCF21 was conditionally deleted in cardiac fibroblasts. Mice were either studied in the absense of injury or after 3 days of myocardial infarction. Cardiac tissue was digested to obtain a single cell suspension and live nucleated cells were sorted for single cell RNA sequencing.
Sample: Tcf21-MCM/fl baseline; scRNA-seq
SAMN40305996 • SRS20699211 • All experiments • All runs
Organism: Mus musculus
Library:
Name: GSM8136445
Instrument: Illumina NovaSeq 6000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC SINGLE CELL
Selection: cDNA
Layout: PAIRED
Construction protocol: Single cells were obtained by flow cytometry of live nucleated cells from cardiac tissue digests. Single-cell droplet libraries were generated according to manufacturers' instructions (Chromium Next GEM Single Cell 3' Reagent Kits v3.1 (Dual index), PN-1000268, 10X Genomics; user guide CG000315). Samples had a greater than 70% viability. About 16,000 cells were loaded to recover 10,000 cells. Briefly, gel emulsion beads (GEM) were generated by combining gel beads, master mix containing the cells and partitioning oil and loaded onto a Chromium Next GEM Chip G at a low dilution. Poly-adenylated mRNA from the cell lysate contained in every GEM was reverse transcribed to cDNA adding an Illumina R1 primer sequence, unique molecular identifier (UMI) and the 10X barcode. The barcoded first-strand cDNA was then purified with MyoOne Silane DynaBeads (20000048, 10x Genomics) and amplified using 14 PCR cycles. The full-length barcoded cDNA was then enzymatically fragmented and size selected using SPRIselect (B23317, Beckman Coulter) and adaptor ligated. The cDNA was then amplified for library construction adding an Illumina R2 primer sequence, paired-end constructs with the P5 and P7 sequences used for Illumina amplification and sample indexes were added.
Runs: 4 runs, 608.8M spots, 84G bases, 24.5Gb
Run# of Spots# of BasesSizePublished
SRR28270765151,618,66020.9G6.1Gb2024-12-03
SRR28270766152,829,55521.1G6.2Gb2024-12-03
SRR28270767152,632,03221.1G6.1Gb2024-12-03
SRR28270768151,724,43920.9G6.1Gb2024-12-03

ID:
32185472

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