ABSTRACT: Primary mice neuronal cultures fractionated through C-18 RP chromatography. Collected fractions analyzed by untargeted LC-MS/MS in positive mode.
Project description:In this project we explore the mitochondrial properties of SCA1+/PDGFRA+/CD31- (S+P+) cardiac fibroblasts (CF) from uninjured adult mouse hearts. We fractionated S+P+ cells on the basis of mitochondrial mass using MitoTracker green-FM and flow cytometry into three fractions: high, mid and low mitochondrial mass as well as total S+P+ cells. Bulk RNA-seq was then performed on each of the fractions.
Project description:The important role of mesenchymal progenitors is quickly emerging in various biological processes. Thus far an appropriate marker has been lacking. Here the fractionation of dissociated skeletal muscle and subsequent enrichment of MPs by an established method was employed. Endothelial and blood derived cells comprised the lineage positive fraction and the remaining cell suspension was fractionated using the surface marker Sca1(Ly6a) to enrich for MPs. RNA sequencing was performed on all fractions and subsequent transcriptomic data was interrogated for MP specific transcription factors. Hic1 was found to be highly enriched in the MP fraction.
Project description:Eukaryotic genomes contain various types of small non-coding RNAs such as microRNAs (miRNAs), silencing RNAs (siRNAs) and PIWI-interacting RNAs (piRNAs). Recent studies point to the presence of a more diverse array of uncharacterized small regulatory RNAs including those that are generated from mRNAs, rRNAs and tRNAs. To explore the possible involvement of tRNA-derived fragments (tRFs) in translational regulation, we fractionated 1 and 8h Drosophila embryos on sucrose density gradients and quantitatively measured by deep-dequencing the expression levels of the tRFs in the fractionated and unfractionated Drosophila embryonic cytosolic extracts. Analysis of 9,007,661 reads has revealed that tRFs, which are produced mainly from the 5’ends of a subset of tRNAs, are mostly associated with the non-polysomal fractions in Drosophila embryos. Quantitative analysis indicates that the expression levels of a subset of tRFs change temporally following the maternal-tozygotic transition in embryos. We detected non-polysomal association of tRFs in S2 cells as well. When transfected into S2 cells, the biotinylated tRFgly:GCC:5 co-fractionated with the non-polysomal complexes similar to the endogenous tRFgly:GCC:5 in embryos and S2 cells. These results suggest that the tRFs, which are much smaller in size than the stres-induced tRNA fragments, are generated selectively from a fraction of tRNAs and that they associate primarily with the non-polysomal complexes in Drosophila embryos and S2 cells.
Project description:Primary human foreskin fibroblasts (HFF) were infected with wild-type simplex virus 1 (HSV-1) strain 17 at a multiplicity of infection (MOI) of 10. Subcellular RNA fractions including total RNA, cytoplasmic RNA, nucleoplasmic RNA and chromatin-associated RNA were prepared and subjected to RNA-seq
Project description:In order to select mRNA transcripts strongly enriched in murine white adipocytes versus brown adipocytes or stromal-vascular fraction, gene expression data of the adipocyte and stromal-vascular fractions of the interscapular brown, inguinal subcutaneous as well as visceral epididymal adipose tissue depots of young adult male C57BL/6 mice housed at constant 23°C ambient temperature were obtained. 18 samples: 3 different adipose tissues separated into stromal-vascular fraction and adipocytes, analyzed in biological triplicates.
Project description:Transcription profile of Escherichia coli cells in biofilms under static batch culture was compared to that of E. coli cells in planktonic cultures. Both E. coli biofilm and planktonic cultures were cultivated for 18 h in 10% Luria-Bertani broth at room temperature (20 degree Celsius). Biofilms were grown in static batch culture in petri dishes. Both planktonic culture and biofilms were homogenized and run through a separated protocol.
Project description:Primary human foreskin fibroblasts (HFF) were infected with the ICP27 null mutant of herpes simplex virus 1 (HSV-1) strain 17 at a multiplicity of infection (MOI) of 10. Subcellular RNA fractions including total RNA, cytoplasmic RNA, nucleoplasmic RNA and chromatin-associated RNA were prepared and subjected to RNA-seq