Project description:The overall data contains expression profile of rat genes of cardiac fibroblasts. G-protein signaling is regulated in cardiac fibroblasts. The expresssion of angiotensin II receptor or cytokine such as interleukin is regulated by PTX (which is suggested to suppress Gi function) treatment. Keywords: control or PTX treatment
Project description:The overall data contains expression profile of rat genes of cardiac fibroblasts. G-protein signaling is regulated in cardiac fibroblasts. The expresssion of angiotensin II receptor or cytokine such as interleukin is regulated by PTX (which is suggested to suppress Gi function) treatment. Keywords: control or PTX treatment Rat cardiac fibroblasts were primarily isolated and plated onto 6 well plate. Cells were treated with water (Ctl 1-3) or PTX for 24 hr (PTX 1-3) and lysed by Buffer RLT. Total RNA was isolated using QIAGEN kit and gene expression was analysed using Affymetrix Gene Chip system and GCOS analysis.
Project description:Gβγ subunits are involved in many different signalling processes in various compartments of the cell, including the nucleus. To gain insight into the functions of nuclear Gβγ, we investigated the functional role of Gβγ signalling in regulation of GPCR-mediated gene expression in primary rat neonatal cardiac fibroblasts. Following activation of the angiotensin II type I receptor in these cells, Gβγ dimers interact with RNA polymerase II (RNAPII). Our findings suggest that Gβ1γ recruitment to RNAPII negatively regulates the fibrotic transcriptional response, which can be overcome by strong fibrotic stimuli. In these specific proteomics experiments, we compared the differential protein abundance in Gβ1 knockdown and WT cardiac fibroblasts following angiotensin II treatment using a bottom-up data-dependent approach.
Project description:We found that cardiac fibroblasts produce and secrete exosomes. miRNA profiling and TaqMan qRT-PCR experiments identified miR-21 expression to be higher in cardiac fibroblasts compared to those of miR-21*, whereas in exosomes miR-21* expression was higher compared to miR-21. The purpose of the study was to validate these findings by miRNA sequencing in cardiac fibroblasts and fibroblasts-derived exosomes. Neonatal rat cardiac fibroblasts were cultured in DMEM + 1% exosome-depleted FBS for 48h. Conditioned medium was collected and exosomes were purified by several centrifugation and filtration steps, following ultracentrifugation. Afterwards total RNA from cardiac fibroblasts and exosomes was isolated for miRNA sequencing.