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MiRNA expression in spatially-sorted hepatocytes

ABSTRACT: miRNA expression in spatially-sorted hepatocytes

PROVIDER: PRJNA556590 | ENA |

REPOSITORIES: ENA

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miRNA expression in spatially-sorted hepatocytes

Project description:The mammalian liver is composed of repeating hexagonal units termed lobules. Spatially-resolved single-cell transcriptomics revealed that about half of hepatocyte genes are differentially expressed across the lobule. Technical limitations impede reconstructing similar global spatial maps of other hepatocyte features. Here, we used zonated surface markers to sort hepatocytes from defined lobule zones with high spatial resolution. We applied transcriptomics, miRNA array measurements and Mass spectrometry proteomics to reconstruct spatial atlases of multiple zonated hepatocyte features. We found that protein zonation largely overlapped mRNA zonation, with the periportal HNF4α as an exception. We identified zonation of miRNAs such as miR-122, and inverse zonation of miRNAs and their hepatocyte target genes, implying potential regulation through zonated mRNA degradation. These targets included the pericentral Wnt receptors Fzd7 and Fzd8 and the periportal Wnt inhibitors Tcf7l1 and Ctnnbip1. Our approach facilitates reconstructing spatial atlases of multiple cellular features in the liver and other structured tissues.

2019-07-26 | GSE134827 | GEO

LC-MS/MS of mouse hepatocytes sorted by lobule layers

Project description:Hepatocytes of the mammalian liver are organized in liver lobules and operate in a spatially-dependent manner. Cells in different positions along the lobule’s porto-cenrtal axis, defined by the directionality of blood flow, express different genes and perform different liver tasks. Gradients of the transcriptome along liver lobule axis has been recently established, yet not for the hepatocyte proteome. We used two surface markers whose levels are inversely zonated – CD73 with a decreasing gradient from pericentral to periportal hepatocytes and E-cadherin with increasing gradient from portal to central hepatocytes. By staining for both surface markers, we efficiently isolated bulk populations of hepatocytes from distinct lobule layers by Fluorescence Activated Cell Sorting (FACS). Over all, we sorted 100,000 hepatocytes from each of eight spatially distinct populations, from five different mice. Cells were washed, digested by trypsin and subjected to LC-MS/MS. More cells from same populations from the same mice were also collected for mRNA sequencing and microRNA microarray profiling, to achieve a multi-omic view on spatially sorted hepatocytes, for better understanding of the transcriptomic and post-transcriptomic levels of regulation of liver zonation.

2019-08-06 | PXD014512 | Pride

Real-time quantitative PCR miRNA analysis of human hepatocytes

Project description:Primary human hepatocytes were treated with compounds modulating steatosis: palmitic acid, compound C and metformin qPCR miRNA expression profiling. Hepatocytes were treated as indicated in the summary. Equal amount total RNA was pooled prior to miRNA expression analysis

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miRNA expression of cultured 4T1 cells sorted by the grade of ALDH activity

Project description:The expression of miRNA was investigated in cultered 4T1 cells sorted by the grade of ALDH activity.

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miRNA expression induced by Sorafenib in HepG2 cells and primary human hepatocytes

Project description:miRNA expression induced by Sorafenib in HepG2 cells and primary human hepatocytes

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Gene expression changes during aging in polyploid and diploid hepatocytes

Project description:To examine gene expression changes duging aging in polyploid and diploid hepatocytes, diploid and polyploid hepatocytes were sorted from young and aged multi-reporter mice and their gene expressions were analyzed by high-throughput RNA sequence.

2021-04-14 | GSE161171 | GEO

MiRNA signatures in cell cycle sorted cells [HeLa_H295R_miRNA_seq]

Project description:For the further examination of cell cycle dependent miRNA expression profile, DNA content based fluorescence activated cell sorting (cell cycle sort) was performed on human transformed cancer cell lines. Phase-dependent miRNA expression profiling was performed on G1, S and G2 phases. Results were validated by quantitative real-time PCR. Phase-dependent miRNA expression profiling was performed on sorted human cancer (cervical - HeLa, adrenocortical - NCI-H295R) cells). G1, S and G2 phases were successfully sorted and analyzed.

2016-05-28 | E-GEOD-73200 | biostudies-arrayexpress

Single-cell sequencing of sorted hepatocytes isolated from livers naïve and LCMV-infected (2 days) C57Bl/6J mice

Project description:Purpose: Elucidate the hepatocyte-specific effects of innate antiviral response to a hepatotropic virus. Methods: Hepatocytes were isolated using a standard 2 step perfusion protocol and viable CD45 negative single cells were sorted on a Sony SH800 instrument und subsequently processed using a 10x Genomics Controller according to standard protocols. Results: Hepatocytes upregulate inflammatory genes and downregulate metabolic genes. Conclusion: Hepatocytes are important innate immune signaling platforms in the liver during viral infection.

2019-12-05 | GSE137082 | GEO

Real-time quantitative PCR miRNA analysis of human hepatocytes

Project description:Real-time quantitative PCR miRNA analysis of human hepatocytes

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MiRNA expression in embryonic murine hepatocytes versus murine cholangiocytes at E18.5

Project description:Differentially expressed miRNAs between hepatocytes (GFP-neg in SOX9-GFP mice) and cholangiocytes (GFP-pos in SOX9-GFP mice) in mice at E18.5. We want to compare miRNA population in biliary cells and parenchymal cells (depleted from hematopoietic cells) during liver development. Cholangiocytes specifically express the transcription factor Sox9. This enables to separate cholangiocytes from the rest of the liver parenchyma by FACS from Sox9-GFP transgenic mice. In conclusion, we want to compare miRNA population in our GFP+ cells (cholangiocytes) and our GFP- cells (liver parenchyma depleted from hematopoietic cells).

2018-12-20 | GSE100145 | GEO

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