Project description:Single Cell Sequencing of FACS purified microglia/myeloid cells isolated with cold mechanical dounce homogenization. Following cell dissociation, the cells from each mouse were split in half and assigned to V3.0 and V3.1 prior to FACS and cell capture.
Project description:Co-housed young adult CEABAC10-transgenic mice (CEA) with C57BL/6NRj background and their wild type littermates (WT) were injected with 1 × 10*4 CFU C. albicans/g body weight via the lateral tail vein (Ca). Control mice received endotoxin-free buffered saline solution (PBS). Mice were sacrificed after 24 h (PBS and Ca) or 72 h (Ca). Peripheral blood was obtained via cardiac puncture. Glycoproteins were captured with hydrazide chemistry and tryptic and PNGase. F-released peptide fractions were analyzed by MS/MS. Quantitative assessment revealed differential glycoprotein expression CEABAC10 mice 72 h post-infection.
Project description:We report the sequencing of total RNA from CD11b+ mouse bone marrow cells in three groups of C57BL/6 mice: 1) Mice treated with three systemic (tail vein) injections (Days 1, 3, and 5) of phosphate buffered saline (PBS) (pH = 7.4), 2) Mice treated with three tail vein injections (Days 1, 3, and 5) of exosomes derived from enzalutamide resistant CWR-R1 prostate cancer cells (EnzR exosomes), and 3) Mice treated with three tail vein injections of EnzR exosomes (Days 1, 3, and 5) and high-density lipoprotein mimetic nanoparticles (HDL NPs) (Days 0, 2, and 4). Mice were sacrificed on Day 6, when mouse bone marrow was harvested and RNA from CD11b+ cells was obtained. We find that mice treated with EnzR exosome exhibit alteration in the expression of the matricellular protein, thrombospondin-1. Moreover, we find that HDL NP pre-treatment prevented thrombospondin-1 downregulation in mice treated with EnzR exosomes. This is consistent with our published findings that HDL NPs inhibit the uptake of EnzR exosomes by CD11b+ mouse bone marrow cells.
Project description:To identify gene expression changes in liver associated with circulating EV that carry high levels of miR-122 (from MDA-MB-231 and MCF10A/miR-122 cells), we analyzed RNA isolated from the liver of PBS- or EV-treated female NOD scid gamma (NSG) mice. Mice had received tail-vein injected EV (or PBS) twice a week for 5 weeks (~10 ug EV per injection). Gene expression in liver from mice treated with EV from MDA-MB-231 or MCF10A/miR-122 cells was compared to cells treated with PBS or EV from MCF10A cells, both of which served as controls in this experiment.
Project description:Control group: injected with normal saline through tail vein; Experimental group: injected with Escherichia coli//Staphylococcus aureus suspension through a tail vein
Project description:Tissue samples from human tonsils and rat pancreas were subjected to a novel laser-based homogenization method, the PIRL-DIVE homogenization, and to a conventional mechanical tissue homogenization procedure (bead mill and a cryo-grinder). The protein species composition of the protein extracts were compared by differential proteome analysis using two-dimensional gel electrophoresis followed by LC-MS/MS analysis and one-dimensional gel electrophoresis (SDS-PAGE) followed by quantitative LC-MS/MS analysis.
Project description:Control group: 250 g adult male rats injected with normal saline through tail vein; Experimental group: 250 g adult male rats injected with E. coli suspension through a tail vein
Project description:To characterize systemic transcriptional changes induced by DHEA treatment, we performed bulk RNA-seq across a panel of tissues collected from control and DHEA-treated mice. Total RNA was extracted from each tissue using a kit-based workflow following mechanical homogenization in lysis buffer. RNA concentration/quality were assessed prior to library construction and high-throughput sequencing.
Project description:Purpose: Traumatic brain injury (TBI) causes 10%–20% of structural epilepsies and 5% of all epilepsies. The lack of prognostic biomarkers for post-traumatic epilepsy (PTE) is a major obstacle to the development of anti-epileptogenic treatments. In this study, we conducted high throughput small RNA-Seq analysis from tail-vein plasma samples collected from a rat model of PTE to discover miRNA biomarker candidates that could serve as prognostic biomarkers for brain damage severity and the development of PTE. Methods: Epileptogenesis was induced in adult male Sprague-Dawley rats by the lateral fluid-percussion-induced TBI. Epilepsy was defined as the occurrence of at least 1 unprovoked seizure during continuous 1-month video-electroencephalography monitoring in the sixth post-TBI month. Small RNA-seq was performed from tail-vein plasma samples collected from a subset of 20 animals (4 sham-operated controls, 7 TBI rats with epilepsy, 9 TBI rats without epilepsy) at 2 days and 9 days post-TBI. RNA was extracted from 200 μl plasma using an miRNeasy Mini Kit. Small RNA-Seq was conducted by GenomeScan (Leiden, the Netherlands). Small RNA library preparation was performed using the Illumina TruSeq Small RNA Sample Prep Kit. Single-end sequencing was performed on the Illumina HiSeq 4000.