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Gene expression of human hematopoietic stem cells (CD34+) stimulated with native & mutant (244) form of myeloid stem-cell transcription factor PU.1 alone & in combination with myeloid specific cytokines (IL3+GMCSF+Flt2+MCSF+SCF)

ABSTRACT: Gene expression of human hematopoietic stem cells (CD34+) stimulated with native & mutant (244) form of myeloid stem-cell transcription factor PU.1 alone & in combination with myeloid specific cytokines (IL3+GMCSF+Flt2+MCSF+SCF)

PROVIDER: PRJNA428367 | ENA |

REPOSITORIES: ENA

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Gene expression of human hematopoietic stem cells (CD34+) stimulated with native & mutant (244) form of myeloid stem-cell transcription factor PU.1 alone & in combination with myeloid specific cytokines (IL3+GMCSF+Flt2+MCSF+SCF)

Project description:Human CD34+ hematopoietic stem cells gives rise to complete blood lineages through the process of hematopoiesis. Here we tried to differentiate a particular lineage i.e myeloid from CD34+ hematopoietic stem cells using a transcription factor PU.1 and its mutated form. Gene expression analysis was performed after treating cells with protein factors and different cytokine combinations. Cells were given six different treatments including control-PU.1 only and PU.1 with cytokines combination; Y244D only and Y244D with cytokines combination and only cytokine combination (IL3, GMCSF, Flt2, MCSF, and SCF). After 5 days of incubation, cells were washed and pellet was preserved in RNAlater (Ambion) to be shipped to Genotypic Technologies Pvt. Ltd. (Bangalore) for microarray analysis.

2020-09-30 | GSE108720 | GEO

Aeromonas hydrophila strain:244

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Nicotinic acetylcholine receptors and EGF induce c-Kit ligand/Stem Cell Factor (SCF) in a β-arrestin-1 and E2F1 dependent manner in NSCLC

Project description:Lung cancer remains the leading cause of cancer-related deaths worldwide. β-arrestin-1 (ARRB1), a scaffolding protein involved in the termination or desensitization of signals arising from activated G-protein-coupled receptors (GPCRs) has been shown to play a role in invasion and proliferation of many cancers, including nicotine-induced proliferation of human non–small cell lung cancers (NSCLCs). In this study, we analyzed nicotine induced and β-arrestin-1 dependent genes from the microarray data. Our analysis show that SCF (Stem cell factor) strongly differentiated smokers from non-smokers implying an important role of this gene in NSCLCs. SCF, a major cytokine is the ligand for the c-Kit proto-oncogene. Here we elucidate the molecular mechanisms by which nicotine as well as EGF induces the expression of SCF in lung adenocarcinoma cell lines A549 and H1650. ChIP assays and transient transfection experiments showed that transcription factor E2F1 can positively regulate SCF expression at the transcriptional level; depletion of E2F1 or β-arrestin-1 prevented the nicotine-mediated induction of SCF. Given that the binding of SCF to c-Kit leads to activation of multiple downstream signaling pathways including Src, PI3K, MEK and EGFR pathways, our data suggest that the SCF plays a central role in lung carcinogenesis, and may be a potential therapeutic target for combating NSCLC. Studies presented here also provide evidence that SCF along with nicotine promotes self-renewal and proliferation of lung cancer stem cells (CSCs). Our findings reveal an important role and prognostic significance of SCF that can serve as a novel prognostic and predictive biomarker for NSCLC.

2013-11-10 | GSE44617 | GEO

Geotrichum candidum strain:LMA-244

Project description:Geotrichum candidum LMA-244 MLST

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Nicotinic acetylcholine receptors and EGF induce c-Kit ligand/Stem Cell Factor (SCF) in a M-NM-2-arrestin-1 and E2F1 dependent manner in NSCLC

Project description:Lung cancer remains the leading cause of cancer-related deaths worldwide. M-NM-2-arrestin-1 (ARRB1), a scaffolding protein involved in the termination or desensitization of signals arising from activated G-protein-coupled receptors (GPCRs) has been shown to play a role in invasion and proliferation of many cancers, including nicotine-induced proliferation of human nonM-bM-^@M-^Ssmall cell lung cancers (NSCLCs). In this study, we analyzed nicotine induced and M-NM-2-arrestin-1 dependent genes from the microarray data. Our analysis show that SCF (Stem cell factor) strongly differentiated smokers from non-smokers implying an important role of this gene in NSCLCs. SCF, a major cytokine is the ligand for the c-Kit proto-oncogene. Here we elucidate the molecular mechanisms by which nicotine as well as EGF induces the expression of SCF in lung adenocarcinoma cell lines A549 and H1650. ChIP assays and transient transfection experiments showed that transcription factor E2F1 can positively regulate SCF expression at the transcriptional level; depletion of E2F1 or M-NM-2-arrestin-1 prevented the nicotine-mediated induction of SCF. Given that the binding of SCF to c-Kit leads to activation of multiple downstream signaling pathways including Src, PI3K, MEK and EGFR pathways, our data suggest that the SCF plays a central role in lung carcinogenesis, and may be a potential therapeutic target for combating NSCLC. Studies presented here also provide evidence that SCF along with nicotine promotes self-renewal and proliferation of lung cancer stem cells (CSCs). Our findings reveal an important role and prognostic significance of SCF that can serve as a novel prognostic and predictive biomarker for NSCLC. Transfection of A549 cell line with control siRNA, Beta arrestin siRNA and then stimulation of nicotine

2013-11-10 | E-GEOD-44617 | biostudies-arrayexpress

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Single-cell RNA sequencing facilitates quantitative analysis of mouse hematopoietic stem/progenitor cells and SCF/2i expanded cells

Project description:Purpose: The goal of this study is to compare the transcriptome profiling of SCF/2i expanded cells to freshly sorted hemtopoietic stem/progenitor cells (HSPCs) at single cell level. Methods: Mouse bone marrow HSPCs (HSC, MEP, CLP, CMP, GMP) and alive SCF/2i expanded cells were sorted by FACS. The FACS-sorted single cell suspensions were washed with ice cold 0.04% BSA in PBS, then loaded onto 3’ library chips as per the manufacturer’s protocol for the Chromium Single Cell 3’ Library (10X Genomics, V2). The 10× Genomics libraries were first sequenced on an Illumina NextSeq 500 aiming at a coverage of 5,000 raw reads per cell to estimate the cell numbers, before being sequenced deeper on an Illumina HiSeq 2500 aiming at a coverage of 50,000 raw reads per cell. (paired-end; read1: 26 cycles; i7 index: 8 cycles; read 2: 98 cycles). Raw sequencing data were pre-processed using the Cell Ranger pipeline (10× Genomics, v 2.1.0). The raw-count datasets from difference cell types were integrated and analyzed by Seurat3 R package. Results: tSNE plot showed that the transcriptome of SCF/2i cells overlapped with that of freshly sorted GMPs. The top 50 highly expressed genes from each comparison were used for analysis by heatmap, which showed that the SCF/2i cells shared a similar gene expression pattern with GMPs, but not other hematopoietic cell lineages. Conclusion: The SCF/2i expanded cells are GMPs.

2021-01-01 | GSE150344 | GEO

Streptomyces sp. KhCrAH-244

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Yersinia pestis strain:244

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Scf-GFP+ cells from the bone marrow and whole bone marrow microarray

Project description:The HSC niche factor SCF is required for HSC maintenance. Using an Scf-GFP knockin mouse, we have identified a perivascular cell type in the bone marrow expressing high level of Scf. To characterize the novel Scf-GFP+ cells from the bone marrow, we performed microarray analysis on these cells.

2012-02-01 | GSE33158 | GEO

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