Transcriptome analysis of luminal and basal cell subpopulations in the lactating versus pregnant mammary gland
Ontology highlight
ABSTRACT: To identify genes specifically expressed in lactating mammary glands, the gene expression profiles of luminal and basal cells from different developmental stages were compared. Comparison of gene expression in luminal and basal cells harvested from the mammary glands of virgin, 18.5 day pregnant and 2 day lactating mice (2 mice per stage).
Project description:To identify genes specifically expressed in lactating mammary glands, the gene expression profiles of luminal and basal cells from different developmental stages were compared.
Project description:To examine Ikaros tumor suppressor mechanisms, we have utilized inducible RNAi to dynamically restore endogenous Ikaros expression in T-ALL driven by its knockdown. This causes rapid transcriptional repression of Notch1 and associated targets including Myc, even in leukemias harboring spontaneous activating Notch1 mutations (producing aberrant ICN1) similar to those found in 60% of human T-ALL. Ikaros restoration results in sustained regression of Notch1-wild type leukemias while endogenous or engineered ICN1 expression promotes rapid disease relapse, indicating that ICN1 functionally antagonizes Ikaros in T-ALL. RNA-seq was performed on T-ALL (Vav-tTA;TRE-GFP-shIkaros primary leukemia ALL211) cells isolated from two untreated and two 3-day Dox-treated mice.
Project description:To examine Ikaros tumor suppressor mechanisms, we have utilized inducible RNAi to dynamically restore endogenous Ikaros expression in T-ALL driven by its knockdown. This causes rapid transcriptional repression of Notch1 and associated targets including Myc, even in leukemias harboring spontaneous activating Notch1 mutations (producing aberrant ICN1) similar to those found in 60% of human T-ALL. Ikaros restoration results in sustained regression of Notch1-wild type leukemias while endogenous or engineered ICN1 expression promotes rapid disease relapse, indicating that ICN1 functionally antagonizes Ikaros in T-ALL. RNA-seq was performed on T-ALL (Vav-tTA;TRE-GFP-shIkaros primary leukemia ALL65) cells isolated from three untreated and three 3-day Dox-treated mice. There were two sequencing runs of each RNA sample.
Project description:To examine Ikaros tumor suppressor mechanisms, we have utilized inducible RNAi to dynamically restore endogenous Ikaros expression in T-ALL driven by its knockdown. This causes rapid transcriptional repression of Notch1 and associated targets including Myc, even in leukemias harboring spontaneous activating Notch1 mutations (producing aberrant ICN1) similar to those found in 60% of human T-ALL. Ikaros restoration results in sustained regression of Notch1-wild type leukemias while endogenous or engineered ICN1 expression promotes rapid disease relapse, indicating that ICN1 functionally antagonizes Ikaros in T-ALL. RNA-seq was performed on T-ALL (Vav-tTA;TRE-GFP-shIkaros primary leukemia ALL101) cells isolated from three untreated and three 3-day Dox-treated mice. There were two sequencing runs of each RNA sample.
Project description:The lactating mammary glands in ruminants play a pivotal economic role in agriculture and food nutrition. The need for an improved 3D in vitro model to study its lactating biology highlights the importance of ongoing research in this field. Overcoming these challenges represents a crucial area of research, with the development of protocols for livestock lactating organoids being a significant goal for the future. In this study, we utilized virgin dairy goats as a model and successfully established a mammary organoid system. These organoids, cultured within an extracellular matrix (ECM) gel, maintained a bilayer structure that closely resembled the original mammary tissue architecture. Importantly, the expression patterns of genes related to fatty acid synthesis and milk proteins in lactating organoids closely mirrored those observed in lactating mammary tissues. Proteomic analysis further confirmed that major milk proteins were highly produced in the lactating organoids. In summary, this study successfully developed a mammary organoid culture system that holds significant promise as a novel platform for cell-based milk production.
Project description:In this study we mapped H3K27me3, H3K4me3 and H3K9me2 marks in three mammary epithelial subsets: MaSC/basal (MS), luminal progenitor (LP) and mature luminal (ML) in the steady-state. In addition, profiles were generated for H3K4me3 and H3K27me3 marks in the MaSC/basal and luminal populations from the glands of ovariectomized or mid-pregnant (12.5 days) mice as well as from control virgin mice. We used ChIPseq analysis to determine histone modification marks in the MS, LP and ML subsets in the steady-state. We then determined the histone modification profiles of MS and sorted luminal (Lum) cells from pregnant (12.5 dP) or ovariectomized (OVX) mice and compared these with the profiles of control virgin mice in order to study the effect of hormones on the mammary epigenomes.
Project description:In this study we mapped H3K27me3, H3K4me3 and H3K9me2 marks in three mammary epithelial subsets: MaSC/basal (MS), luminal progenitor (LP) and mature luminal (ML) in the steady-state. In addition, profiles were generated for H3K4me3 and H3K27me3 marks in the MaSC/basal and luminal populations from the glands of ovariectomized or mid-pregnant (12.5 days) mice as well as from control virgin mice.
Project description:Triplicate RNA-seq expression analysis of bone marrow pre-B cells isolated from mice, to demonstrate repertoire at the IgH locus Triplicate RNA-seq expression analysis of bone marrow pre-B cells
Project description:Hypomorphic mutations of PAX5 occur in one third of B-progenitor acute lymphoblastic leukemias (B-ALLs), however their functional consequences remain undefined. Here we employ advanced transgenic RNAi in mice to suppress endogenous Pax5 expression in the hematopoietic compartment in vivo, which co-operates with activated STAT5 to induce B-ALL. In this model, restoring endogenous Pax5 expression in established B-ALL induces transcriptional and immunophenotypic changes reminiscent of normal B cell differentiation, disabling leukemia-initiating capacity and ultimately causing leukemia clearance. Comparison of leukemias harvested from triplicate untreated mice versus triplicate Dox-treated (3 days) mice
Project description:Toxoplasma gondii is an obligate intracellular Apicomplexan parasite capable of invading and surviving within nucleated cells in most warm-blooded animals. This remarkable task is achieved through the delivery of effector proteins from the parasite into the parasitophorous vacuole and host cell cytosol that rewire host cellular pathways, facilitating parasite evasion of the immune system. Here, we have identified a novel export pathway in Toxoplasma that involves cleavage of effector proteins by the Golgi-resident aspartyl protease 5 (ASP5) prior to translocation into the host cell. We demonstrate that ASP5 cleaves a highly constrained amino acid motif that has some similarity to the PEXEL motif of Plasmodium parasites. We show that ASP5 can mature effectors at both the N- and C-terminal ends of proteins and is also required for the trafficking of proteins without this motif. Furthermore, we show that ASP5 controls establishment of the nanotubular network and is required for the efficient recruitment of host mitochondria to the parasitophorous vacuole membrane. Global assessment of host gene expression following infection reveals that ASP5-dependent pathways influence thousands of the transcriptional changes that Toxoplasma imparts on its host cell. This work characterizes the first identified machinery required for export of Toxoplasma effectors into the infected host cell. Three groups of human foreskin fibroblasts are compared. Each group has 3 replicates giving a total of 9 samples. The first group of samples are infected with wild type (GRA16HA) Toxoplasma gondii, the second group with Asp5 knock-out Toxoplasma gondii, and the final group remain uninfected. All fibroblasts are generated from one donor sample.