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Glucocorticoid receptor ChIP-seq in an immortalized podocyte cell line

ABSTRACT: Glucocorticoid receptor ChIP-seq in an immortalized podocyte cell line

PROVIDER: PRJEB17855 | ENA |

REPOSITORIES: ENA

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Glucocorticoid receptor ChIP-seq in an immortalized podocyte cell line

Project description:The identification of the glucocorticoid receptor cistrome in a conditionally immortalized human podocyte cell line developed by transfection with the temperature-sensitive SV40-T gene

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Gene expression in a conditionally immortalized mouse podocyte cell line

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Glucocorticoid therapy regulates podocyte motility by inhibition of Rac1

Project description:Nephrotic syndrome (NS) occurs when the glomerular filtration barrier becomes excessively permeable leading to massive proteinuria. In childhood NS, dysregulation of the immune system has been implicated and increasing evidence points to the central role of podocytes in the pathogenesis. Children with NS are typically treated with an empiric course of glucocorticoid (Gc) therapy; a class of steroids that are activating ligands for the glucocorticoid receptor (GR) transcription factor. Although Gc-therapy has been the cornerstone of NS management for decades, the mechanism of action, and target cell, remain poorly understood. We tested the hypothesis that Gc acts directly on the podocyte to produce clinically useful effects without involvement of the immune system. In human podocytes, we demonstrated that the basic GR-signalling mechanism is intact and that Gc induced an increase in podocyte barrier function. To gain mechanistic insight we performed RNA microarray and ChIP-sequencing and identified Gc regulation of motility genes.

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Project description:The change of mRNA expression in murine immortalized podocyte were analyzed after miR-26a silencing. These results provide a basical information of molecular pathology in podocyte biology. Mouse podocytes immortalized by temperature sensitive SV40 were used. Podocyte cultures grown at 33 °C were trypsinized and then cultured with RPMI-1640 without antibiotics in 24-well plates at 60–70% confluence for 2 days. On day 3, an anti-miR negative control (40 pmol) or the miR-26a miRNA inhibitor (40 pmol) was transfected to podocytes. The cells were analyzed after culturing for 24 hour.

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RNAseq in immortalized human podocyte cell line following PRDM15 CRISPR knock-out

Project description:We recently identified variants in PRDM15 in affected individuals with proteinuric kidney disease. To determine the downstream pathways mediated by PRDM15 knock-down, PRDM15 knock-out immortalized podocyte cell lines were generated using CRISPR/Cas genome editing. Two different guide RNAs were utilized to generate two different cell lines, while a scramble guide RNA was utilized as control. We performed RNAseq and identified 151 differentially regulated genes, which coalesced upon pathways involved in renal development.

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Genome-wide profiling of Glucocorticoid receptor binding in A549, Nalm-6, MEF WT and MEF GCN5/PCAF double knockout (ChIP-seq)

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RNAseq in immortalized human podocyte cell line following PRDM15 CRISPR knock-out

Project description:RNAseq in immortalized human podocyte cell line following PRDM15 CRISPR knock-out

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