Project description:To analyse the transcriptome landscape of differentiating cardiomyocytes, GFP labeled cardiac cells were purified by fluorescent activated cell sorting using the TinC*>GFP transgenic line, and expression profile analysed by deep sequencing.
Project description:Background Proteomic characterization of microglia has been limited by low yield and contamination by non-microglial proteins by magnetic-activated cell sorting (MACS) enrichment strategies. To determine whether a fluorescent-activated cell sorting (FACS)-based strategy overcomes these limitations, we compared microglial proteomes of MACS and FACS-based purified CD11b+ microglia in order to identify core sets of microglial proteins in adult mouse brain tissue. Results Quantitative multiplex proteomics by tandem mass tag mass spectrometry (TMT-MS) of MACS-enriched (N = 5) and FACS-purified (N = 5) adult wild-type CD11b+ microglia identified 1,791 proteins, of which 953 were differentially expressed indicating significant differences between both approaches. While the FACS-purified microglia proteome was enriched with cytosolic, endoplasmic reticulum and ribosomal proteins involved in protein metabolism and immune system functions, the MACS-enriched microglia proteome was enriched with proteins related to mitochondrial function and synaptic transmission. As compared to MACS, the FACS microglial proteome showed strong enrichment for canonical microglial proteins while neuron, astrocyte, and oligodendrocyte proteins were decreased. Interestingly, we observed enrichment of endothelial specific proteins in the FACS microglia proteome. By comparing FACS-purified microglia proteomes with transcriptomes, we observed highly concordant as well as highly discordant proteins that were abundant at the protein level but low at the transcript level. Conclusions We demonstrate that TMT-MS proteomics of FACS-purified adult microglia is superior to column-based enrichment approaches, resulting in purer and highly-enriched microglial proteomes. We also define core sets of highly-abundant adult microglial proteins including Moesin (Msn) that can guide future studies.
Project description:Differences of metabolism-related gene expression profiles in human ESCs and ESC-derived purified cardiomyocytes were analyzed and successfully identified. Human ESCs and ESC-derived purified cardiomyocytes were used for this experiment.
Project description:This SuperSeries is composed of the following subset Series:; GSE2039: FACS purified cortical projection neurons; GSE17783: Analysis of gene expression in FACS-purified cortical projection neurons using Affymetrix 430 2.0 microarrays Experiment Overall Design: Refer to individual Series
Project description:3 subtypes of cortical projection neurons were purified by fluorescence-activated cell sorting at 4 different stages of development from mouse cortex. A detailed description of the data set is described in Arlotta, P et al (2005). Keywords = corticospinal motor neuron callosal corticotectal cortex development FACS
Project description:We used microarrays to profile global gene expression changes of Pou5f1-GFP-positive germ cells between E11.5 to E15.5. Germ cells were FACS-purified from gonadal single cell suspension based on Pou5f1-GFP expression. Three timepoints were included in this study: E11.5 (male/female), E13.5 (male) and E15.5 (male). For each timepoint, three biological replicates were analyzed. The Pou5f1-GFP-negative (non-germ cell) fraction of E13.5 (male) gonads was also included as a control.
Project description:NK cells from NKDxIL15tg mice spleens and bone marrow were purified by FACS. NK cells from IL15tg mice spleens were purified by FACS. Gene expression differences were analyzed to identify genes involved in NK cell development. Experiment Overall Design: Immature NK cells from NKDxIL15tg mice and mature NK cells from IL15tg mice were purified by FACS. RNA was made for target synthesis and hybridization to Affymetrix microarrays.
Project description:We sequenced mRNA from HFSCs that were FACS purified (CD34+alpha-6 Integrin+ cells) from p21 mouse epidermis, total epidermal cells from p21 mice and HFSCs cultured for 14 days in 3C cultures, 3 biological replicates each. We sequenced mRNA from different cell populations that were FACS-purified from HFSC cultures (also known as 3C cultures), 3 biological replicates each. We performed two independent comparisons: 1. Comparison of FACS-purified cells grown in standard 3C cultures: HFSCs (CD34+ alpha-6 Integrin+) and non-HFSCs (CD34neg alpha-6 Integrin+). 2. Comparison of total cells in 3C cultures treated for 48 hours with dorsomorphin (BMP inhibitor that causes depletion of CD34+ alpha-6 integrin+ cells in 3C cultures) or with cyclopamine Shh inhibitor that causes enrichment of CD34+ alpha-6 integrin+ cells in 3C cultures). Overall design: Examination of transcriptomes from bona fide HFSCs, total epidermal cells and cultured HFSCs. The aim of the experiment was to determine: for the first comparison (1.) the identity of the two populations of cells that grow in 3C cultures (CD34+ alpha-6 Integrin+ and CD34neg alpha-6 Integrin+); and for the second comparison (2.) the early transcriptional changes that lead to a fate change in cells grown in 3C cultutres to drive depletion (with dorsomorphin treatment) or enrichment (with cyclopamine treatment) of HFSCs.
Project description:3 subtypes of cortical projection neurons were purified by fluorescence-activated cell sorting (FACS) at 4 different stages of development from mouse cortex. A detailed description of the data set is described in Arlotta, P et al (2005) and Molyneaux, BJ et al (2009). The hybridization cocktails used here were originally applied to the Affymetrix mouse 430A arrays and submitted as GEO accession number GSE2039. The same hybridization cocktails were then applied to the Affymetrix mouse 430 2.0 arrays, and those data are contained in this series. Experiment Overall Design: Three subtypes of cortical neurons were purified by FACS at multiple stages of mouse brain development. The neuron subtypes are: corticospinal motor neurons (CSMN), callosal projection neurons (CPN), and corticotectal projection neurons (CTPN). The stages of development included embryonic day 18 (E18), postnatal day 3 (P3), postnatal day 6 (P6), and postnatal day 14 (P14). CSMN and CPN were analyzed at all four stages, while CTPN were only analyzed at P14. The replicates included in the data set are all true biological replicates with independent sample collection for each.