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Expression data from a mouse small cell lung cancer (SCLC) cell line, KP1, transfected with Notch-ICD

ABSTRACT: Transient transfection of activated Notch1 (Notch1-ICD) decreases cellular proliferation and reduces the expression of a subset of neuroendocrine genes. We used microarrays to identify the gene expression changes 48 hours after overexpression of Notch1-ICD. mSCLC cells transfected with Notch1-ICD-ires-GFP were FACS sorted for GFP+ cells 48 hours after transfection. RNA was then extracted and hybridized to Affymetrix GeneChip® Mouse Gene 2.0 ST array.

ORGANISM(S): Mus musculus

SUBMITTER: Julien Sage

PROVIDER: E-GEOD-69091 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Expression data from a mouse small cell lung cancer (SCLC) cell line, KP1, transfected with Notch-ICD

Project description:Transient transfection of activated Notch1 (Notch1-ICD) decreases cellular proliferation and reduces the expression of a subset of neuroendocrine genes. We used microarrays to identify the gene expression changes 48 hours after overexpression of Notch1-ICD.

2015-05-21 | GSE69091 | GEO

Gene Expression Profiling on Stably Expressing ICD JM-a CYT-1 or CYT-2 ERBB4 isoforms in MCF10A cells

Project description:Convergent and divergent effects of two ICD ERBB4 isoforms on gene expression in normal-like MCF10A mammary epithelial cells lacking endogeneous ERBB4 expression. Control vector (VEC) and ERBB4 expression MCF10A cells (ICD CYT1 or CYT2) maintained in regular medium containing 10% serum, were collected and RNA samples were collected and analyzed.

2014-05-07 | E-GEOD-57339 | biostudies-arrayexpress

Perturbation of the Notch pathway in Xenopus laevis embryos

Project description:The Notch signaling pathway functions in a number of processes during embryologic development, especially the maintenance or aquisition of cell fate. We purturb the Notch signalling pathway in embryonic Xenopus laevis in order to 1) better characterize the downstream targets of Notch signalling, and 2) determine the extent to which early embryos can recover from transient purturbations to critical signalling pathways, if at all. Xenopus laevis embryos were unilaterally injected at the two cell stage with either GFP, GFP and ICD (Notch intracellular domain, an up-regulator of the Notch pathway), or GFP and DBM (domain-binding mutant, a downregulator of the Notch pathway). At stages 18, 28, and 38, for each injection, pooled total RNA from 10 embryos was extracted. Extraction was performed for three biological replicates for each time/injection condition. cDNA from total RNA was hybridized on Affymetrix Xenopus laevis Genome 2.0 arrays.

2014-08-06 | E-GEOD-60127 | biostudies-arrayexpress

Comparing the effects of MITF on transcriptional regulation to the TEN1-ICD

Project description:We identified histidine triad nucleotide binding protein 1 (HINT1) as a human teneurin-1 ICD interaction partner in a yeast-2 hybrid screen. This interaction was confirmed in human cells, where HINT1 is known to inhibit the transcription of target genes by directly binding to transcription factors at the promoter. In a whole transcriptome analysis of BS149 glioblastoma cells overexpressing the teneurin-1 ICD, several microphthalmia-associated transcription factor (MITF) target genes were found to be up-regulated. Interestingly, MITF is one of the transcription factors inhibited by HINT1. Thus, we directly compare the transcriptomes of MITF versus TEN1-ICD overexpressing BS149 cells in this study, in order to reveal any co-regulated genes. For the whole transcriptome analysis of MITF, cells were transiently transfected in triplicates with either, pcDNA3.1-RFP-HA (negative control) or pcDNA3.1-MITF-RFP-HA. The overexpressing cells were then FACS-sorted directly into RLT lysis buffer (Qiagen) at a 3:1 volume ratio of lysis buffer to cells in PBS, 24 h post-transfection.

2015-02-04 | E-GEOD-61705 | biostudies-arrayexpress

Studying the effects of the TEN1-ICD on transcriptional regulation

Project description:Teneurins are large type II transmembrane proteins that are necessary for the normal development of the central nervous system (CNS). While many studies highlight the significance of teneurins, especially during development, there is only limited information known about the molecular mechanisms of function. Previous studies have shown that the N-terminal intracellular domain (ICD) of teneurins can be cleaved at the membrane and subsequently translocates to the nucleus where it can influence gene transcription. Target genes as well as mechanisms have yet to be elucidated, and thus we are investigating the transcriptional activity of the human teneurin-1 ICD in this study. For the whole transcriptome analysis of TEN1-ICD overexpression, we used a modified and improved tet-system. Two separate vectors are required to make the cell line stable. One contains the tet-activating domain fused to a glucocorticoid binding domain (GBD). The other contains the tetO operator sequences directly upstream of a CMV promoter and the gene to be overexpressed. BS149 cells were first transfected with pirtetR-GBD and made stable by Puromycin selection, and then after further transfection with either ptetO-eGFP-His (negative control) or ptetO-TEN1-ICD-eGFP-His by Hygromycin selection. The stable BS149 cell lines were split into three 10 cm Petri dishes each. The triplicate cell lines were cultured once before induction with Dexamethasone and Doxycycline. The overexpressing cells were then FACS-sorted directly into RLT lysis buffer (Qiagen) at a 3:1 volume ratio of lysis buffer to cells in PBS, 24 h post-induction.

2015-02-04 | E-GEOD-61704 | biostudies-arrayexpress

Gene expression in mouse pre-B cells transduced with Ikaros.

Project description:Ikaros family DNA binding proteins are critical regulators of B cell development. To identify Ikaros-regulated genes in pre-B cells we performed gene expression studies at enhanced temporal resolution. We used retroviral gene transfer to express Ikaros proteins in the pre-B cell line B3. Total RNA from 2 biological replicates of B3 cells transduced with wild type Ikaros (HA-Ikaros-IRES-GFP) and DNA binding-deficient Ikaros mutant 159A (HA-159A Ikaros-IRES-GFP) was isolated 48h after infection. To increase the temporal resolution of Ikaros-regulated gene expression we transduced B3 cells with inducible Ikaros (HA-Ikaros-ERt2-IRES-GFP) and isolated total RNA from 3 biological replicates after 2 h and 6 h of exposure to 4-hydroxytamoxifen. Vector transduced B3 cells (ERt2-IRES-GFP) treated with 4-hydroxytamoxifen were used as controls.

2013-01-15 | E-GEOD-38110 | biostudies-arrayexpress

Effects of TrkB-ICD overexpression on the morphology, activity and transcriptome of cultured primary cortical neurons

Project description:In Alzheimer’s disease (AD), amyloid β (Aβ)-triggered cleavage of TrkB-FL impairs brain-derived neurotrophic factor (BDNF) signaling, thereby compromising neuronal survival, differentiation, as well as synaptic transmission and plasticity. In addition to compromising canonical BDNF signalling pathways, TrkB-FL cleavage produces an intracellular fragment (TrkB-ICD), which was shown to accumulate in the nucleus and to display tyrosine kinase activity. To dissect the role of TrkB-ICD overexpression from the loss of endogenous signaling throught TrkB-FL, we used lentiviruses to overexpress the TrkB-ICD sequence in cultured primary cortical neurons and performed morphological, electrophysiological and transcriptomics studies. While TrkB-ICD overexpression did not affect cell survival, it caused a significant decrease in the number of dendritic spines, both compared to untransduced and GFP-transduced neurons. Furthermore, TrkB-ICD overexpressing neurons presented a hyperpolarized resting membrane potential and increased frequency of miniature excitatory postsynaptic currents (mEPSCs). Finally, TrkB-ICD overexpression was associated with the upregulation of genes involved in (i) neuronal survival, growth and differentiation; (ii) neuronal cytoarchitecture and spine morphology; (iii) neurodegenerative processes, including AD; and (iv) synaptic transmission and plasticity. Overall, these results show that TrkB-ICD overexpression causes dendritic spine loss, alters excitatory synaptic transmission and causes transcriptome-wide changes, namely in genes coding for proteins involved in synaptic processes.

2024-07-30 | GSE271487 | GEO

microarray_submission

Project description:We evalueted gene expression in dorsal root ganglion (DRG) over-expressing NRG1 typeIII ICD (Intra Cellular Domain). DRG were infected with a lentivirus expressing ICD and compared to not infected DRG or DRG infected with a lentivirus expressing EGFP.

2014-12-14 | E-MTAB-2131 | biostudies-arrayexpress

RNA-sequencing comparison of adipose tissue endothelial cells with Notch1 ICD overexpression

Project description:RNA-sequencing comparison of adipose tissue endothelial cells with Notch1 ICD overexpression

| PRJNA878482 | ENA

Analysis of CITED4 overexpression in the CRC cell line SW480

Project description:Ires-GFP and CITED4-ires-GFP overexpressing permanent cell lines were generated by transfection of the CRC cell line SW480 with either the ires-GFP containing vector vector (in pCDNA4HISMAX) or the CITED4-ires GFP in (pCDNA4HISMAX) and selected with 2 ug/ ml puromycin in order to prepare permanent cell lines. For microarray analysis, the two cell lines were plated at a density of 1 x 10E5 cell in 12 well plates in 1 ml RPMI plus 10% FCS and penicillin/ streptomycin. The respective cell lines were harvested at day 1, 3 and 5 for RNA preparation and microarray analysis using Agilent human 4 x 44k expression microarrays. Every time point (cy3 labeled) was hybridzed together with Stratagene Human Total RNA standard as a control (cy5). Comparisons were made between the CITED4 overexpressing permanent cell line and ires-GFP containing cell line.

2014-11-01 | E-MTAB-2486 | biostudies-arrayexpress

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