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Interaction effects of knockdown of PML and TNFalpha treatment in HUVEC cells

ABSTRACT: To study the interaction effects between promyelocytic leukemia (PML) gene knockdown and tumor necrosis factor-alpha (TNFalpha) signaling in human umbilical vein endothelial cells (HUVECs), we transfected control or two independent PML siRNAs into HUVEC cells without or with 20 ng/mL TNFalpha treatment for 20 h. The total RNA was extracted for gene expression microarray analyses.

ORGANISM(S): Homo sapiens

PROVIDER: GSE35000 | GEO | 2013/01/10

SECONDARY ACCESSION(S): PRJNA151067

REPOSITORIES: GEO

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Interaction effects of knockdown of PML and TNFalpha treatment in HUVEC cells

Project description:To study the interaction effects between promyelocytic leukemia (PML) gene knockdown and tumor necrosis factor-alpha (TNFalpha) signaling in human umbilical vein endothelial cells (HUVECs), we transfected control or two independent PML siRNAs into HUVEC cells without or with 20 ng/mL TNFalpha treatment for 20 h. The total RNA was extracted for gene expression microarray analyses. The experiment is a 3x2 two-factor design. One factor is siRNA and it has three levels (siCtrl, siPML1, siPML2). siCtrl is RISC-inducing non-targeting control siRNA. siPML1 and siPML2 designate two independent siRNAs against the PML gene. Two independent siRNAs were used to eliminate the off-target effects of siRNA. The other factor is TNFalpha treatment (20 ng/mL for 20 h) and it has two levels: Untreated (U) or Treated (T). Each sample has technical duplicates.

2013-01-10 | E-GEOD-35000 | biostudies-arrayexpress

The effects of knockdown of PML by different individual siRNAs in HUVEC cells

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RNA sequencing of HECTD3 knockdown during TNFalpha stimulation in primary HUVEC cells

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The effects of knockdown of PML by different individual siRNAs in HUVEC cells

Project description:To study the potential target genes regulated by PML in endothelial cells, we carried out siRNA-mediated knockdown of PML in HUVEC cells. To eliminate the off-target effects of siRNAs, we utilized two different siRNAs. Only the genes changed in the same pattern following both siRNAs transfection are considered as potential PML-knockdown responsive genes. The experiment is one-factor (siRNA) with three ranks (siCtrl, siPML-1, siPML-2). We included technical duplicates for each sample. To minimize systemic errors assoicated with technique, we distributed the duplicates on two different microarray chips.

2012-05-30 | E-GEOD-29238 | biostudies-arrayexpress

PML regulates NF-ÎºB phosphorylation and transcriptional activity

Project description:The transcription factor NF-ÎºB is considered the master regulator of the immune response but also acts broadly to regulate gene expression that influences cell survival, proliferation and differentiation. Post-translational modification of NF-ÎºB, phosphorylation in particular, is essential for the transactivation activity of NF-ÎºB. Emerging evidence suggests that the regulation of NF-ÎºB in the nucleus is critical in controlling gene expression. Promyelocytic Leukemia (PML) is a nuclear protein that forms nuclear bodies (PML NBs), sub-nuclear structures that are associated with transcriptionally active genomic regions that have been implicated in multiple processes such as apoptosis, senescence and anti-viral responses. Chromosomal translocations leading to the expression of a PML-retinoic acid receptor-Î± (PML-RARÎ±) fusion protein are causative for acute promyelocytic leukemia (APL) characterised by a differentiation block at the promyelocytic state of myeloid development. Here we demonstrate that PML is required for phosphorylation of NF-ÎºB p65 and that PML is essential for NF-ÎºB- induced transcriptional responses. Our analysis of available transcriptional profiles of all-trans retinoic acid treated acute promyelocytic leukemia (APL) cells identifies a NF-ÎºB transcriptional programme suppressed by PML-RARÎ±. We further demonstrate that PML-RARÎ± inhibits NF-ÎºB phosphorylation and transcriptional activity. Our findings demonstrate a critical role for PML in promoting NF-ÎºB transcriptional activity which may contribute to APL initiation and maintenance. WT and PML-/- MEFs were analysed for gene expression analysis. Total of 12 samples, inlcluding triplicates were utilized. WT MEFs and PML-/- were stimulated with TNFÎ± for three hours and analysed for gene expresison using unstimulated WT MEFs as control.

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Profiling of PML-body associated regions

Project description:The promyelocytic leukemia (PML) body is a phase-separated nuclear structure composed of various proteins including several chromatin regulators, and physically associates with chromatin. To address functional roles of the PML-chromatin association, we conducted genome-wide profiling of PML body-associated regions.

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Expression profiling in Human Umbilical Vein Endothelial Cells (HUVEC)

Project description:Whole transcriptome gene expression profiling of Normal -(HUVEC) Human Umbilical Vein Endothelial Cells The HUVEC gene expression results analyzed in this study are further described in Kustermann S. et al. (2014) A real-time impedance based screening assay for drug induced vascular leakage. A NimbleGen Homo sapiens Expression Array [100718_HG18_opt_expr] study using total RNA recovered from HUVECs grown until 80% confluency. Each microarray measures the expression level of 23,611 genes using 45,033 probe sets with three 60-mer probes (PM) per probe set. Each probe set is represented once on the array.

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RNA sequencing of HECTD3 knockdown during TNFalpha stimulation in primary HUVEC cells

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