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The gene expression profile of primary BMMs differentiating into osteoclasts after knockout of solute carrier family 11 member 2 (SLC11A2)

ABSTRACT: To investigate the effect of solute carrier family 11 member 2 (SLC11A2) knockout on osteoclast differentiation, BMMs were extracted from the femurs of SLC11A2 knockout mice and the same litter of control mice, treated with RANKL, and then rna sequencing was performed.

ORGANISM(S): Mus musculus

PROVIDER: GSE283491 | GEO | 2024/12/15

REPOSITORIES: GEO

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The gene expression profile of primary BMMs differentiating into osteoclasts after knockout of solute carrier family 11 member 2 (SLC11A2)

Project description:The gene expression profile of primary BMMs differentiating into osteoclasts after knockout of solute carrier family 11 member 2 (SLC11A2)

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High-throughput sequencing analysis of BMMs and BMMs-derived osteoclasts

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Project description:To profile the overall transcriptomic changes, we performed RNA-seq with bone marrow macrophages (BMM) before and after osteoclast differentiation (day 0 vs day 4). To induce mature osteoclasts formation, BMM were seeded and culture with customized αMEM containing 1/50 CMG14-12 supernatant and 75 ng/ml RANKL for 4 days.

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Comparative characterization of osteoclasts derived from murine bone marrow macrophages and RAW 264.7 cells using quantitative proteomics

Project description:Osteoclasts (OCs) are bone-resorbing cells differentiated from macrophage/monocyte precursors in response to M-CSF and RANKL. In vitro models are principally based on primary bone marrow macrophages, but RAW 264.7 cells are frequently used because they are widely available, easy to culture, and more amenable to genetic manipulation than primary cells. Increasing evidence, however, has shown that the vastly different origins of these two cell types may have important effects on cell behavior. In particular, M-CSF is prerequisite for the differentiation of BMMs, by promoting survival and proliferation and priming the cells for RANKL induction. RAW 264.7 cells readily form OCs in the presence of RANKL, but M-CSF is not required. Based on these key differences, we sought to understand their functional implications and how it might affect osteoclast differentiation and related signaling pathways. Using a robust and high-throughput proteomics strategy, we quantified the global protein changes in OCs derived from bone marrow macrophages and RAW 264.7 cells at 1, 3, and 5 days of differentiation. Correlation analysis of the proteomes demonstrated low concordance between the two cell types (R2 ≈ 0.13). Bioinformatics analysis indicate that RANKL-dependent signaling was intact in RAW 264.7 cells, but biological processes known to be dependent on M-CSF were significantly different; including cell cycle control, cytoskeletal organization, and apoptosis. RAW 264.7 cells exhibited constitutive activation of Erk and Akt that was dependent on the activity of Abelson tyrosine kinase, and the timing of Erk and Akt activation was significantly different between BMMs and RAW 264.7 cells. Our findings provide the first evidence for major differences between BMMs and RAW 264.7 cells, indicating that careful consideration is needed when using the RAW 264.7 cell line when studying M-CSF-dependent signaling and functions.

2018-11-27 | PXD009610 | Pride

Effect of NVP-BHG712 treatment on gene expression of BMMs differentiating into osteoclasts.

Project description:To investigate the effect of NVP-BHG712 on osteoclast differentiation, BMMs were removed from femorae of C57BL/6 mice and treated with NVP-BHG712 for RNA-seq

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Solute carrier family 12 member 5 promotes tumor development of intrahepatic cholangiocarcinoma

Project description:Intrahepatic cholangiocarcinoma (ICC) is the second most common primary malignant tumor of the liver, characterized by high malignancy, poor prognosis, and lack of effective treatment.Solute carrier family 12 member 5 (SLC12A5) is frequently overexpressed in many tumors and is involved in the development and progression of several human cancers. Here, we aimed to investigate the expression and biological function of SLC12A5 in ICC.

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Microarray studies on genes stimulated by RANKL in bone marrow macrophages in a cyclooxygenase-2-dependent manner

Project description:Overall goal was to look for the differentially expressed genes between receptor activator of nuclear factor kappa-B ligand (RANKL)-stimulated wild type (WT) bone marrow macrophages (BMMs) and the BMMs in which the expression or the enzyme activity of cyclooxygenase-2 (Cox2), the major enzyme for prostaglandin E2 (PGE2) synthesis, is absent. For this purpose, two separate microarrays (experiment 1 and 2) were done. In both the experiments, Serum Amyloid A3 (Saa3) was one of the most highly differentially expressed gene. For experiment 1, BMMs were isolated from the bone marrow of Cox2 knockout (KO) and WT mice. Out of the three sample groups, two were treated with 30 ng/ml each of macrophage-colony stimulating factor (M-CSF) and RANKL, while the third group was treated with M-CSF only, for 1 day. Two of the sample groups had n=3 biological replicates (i.e. samples), while the third one had n=4 biological replicates. The differential gene expression comparisons among the sample groups were done as (A) WT+MCSF+RANKL_d1 versus Cox2KO+MCSF+RANKL_d1 and (B) WT+MCSF+RANKL_d1 versus WT+MCSF_d1. For experiment 2 also, BMMs were obtained from Cox2 KO and WT mice. All the four sample groups were treated with M-CSF and RANKL for 3 days. All the four sample groups had n=4 biological replicates. One of the WT sample group was also treated with an inhibitor of Cox2 activity,NS398 (0.1 Âµm) and one of the KO sample group was also treated with the product of Cox2 enzyme, PGE2 (1 Âµm). The differential gene expression comparisons among the sample groups were done as (A) WT+MCSF+RANKL_d3 versus Cox2KO+MCSF+RANKL_d3; (B) WT+MCSF+RANKL_d3 versus WT+MCSF+RANKL+NS398_d3 and (C) Cox2KO+MCSF+RANKL+PGE2_d3 versus Cox2KO+MCSF+RANKL_d3.

2016-01-07 | E-GEOD-72047 | biostudies-arrayexpress

Gene expression microarray data of bone marrow derived macrophages using GM-CSF(GM-BMMs) co-cultured with E0771 breast tumor cells

Project description:E-FABP is dominately expressed in macrophages/dendritic cells. Thus E-FABP may play a siginificant role in their immune response to tumor insult. We treated mouse GM-BMMs of different genotype with E0771 breast cancer cells. Then we investigate the global gene expression in these GM-BMMs in response to tumor treatment. Bone marrow cells were collected from naïve C57BL/6 and E-FABP knockout mice of 6~8 week old. GM-BMMs were derived using GM-CSF (in RPMI1640 with 5%FBS) for 7 days before tumor treatment. After 18hr tumor teatment, GM-BMM total RNA was extracted using RNeasy Mini Kit (Qiagen) and subjected to mouse mRNA Gene expression by Affymetrix microarray.

2014-09-02 | E-GEOD-54073 | biostudies-arrayexpress

Murine Bone Marrow Macrophages: Control vs. RANKL Stimulated

Project description:To identify the microRNAs that are involved in osteoclastogenesis, microRNA expression profiles in mouse bone marrow macrophages (BMMs) stimulated with RANKL (BMOc) were compared with that of control untreated BMMs. These results provide insights into the mechanisms to regulate osteoclastogenesis and bone resorption activities in osteoclasts by microRNA. BMMs were cultured with 20 ng/ml M-CSF in the presence or absence of 50 ng/ml RANKL for 24 hours. Cells were collected for total RNA isolation, and were subjected to microRNA array analysis.

2013-07-10 | E-GEOD-48629 | biostudies-arrayexpress

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