ABSTRACT: Comprehensive in vitro expression profiling of small chemical compound inhibitors of TGFbR1 kinase activity. Additional file E-MTAB-265.additonal.1.zip contains excel file with Taqman data validating the array normalization method
Project description:The investigation of the function of SUPT6H, SF3B1 and HNRNPH1 in Ewing sarcoma The expression profiles of TC32 Ewing sarcoma cells were determine 48 hours post siRNA-transfected. Groups consisted of untreated cells (three samples), and negative control (siNeg) transfected-TC32 cells (six samples), and TC32 cells in which either SUPT6H, SF3B1 or HNRNPH1 were silenced; three different siRNAs corresponding to each gene were assessed separately.
Project description:This microarray was performed to gain insight in the downstream targets of GY118F in iPS cells. This array is part of the following paper to be published in Nature Communications: “JAK/STAT3 signalling is sufficient and dominant over antagonistic cues for the establishment of naïve pluripotency” by Anouk L. van Oosten, Yael Costa, Austin Smith & José C.R. Silva The samples analysed were: - GY118F iPS cells derived and maintained in N2B27 plus GCSF (t0, indicated in data as nlng t0 a), -same cells but from which GCSF was withdrawn for 12 or 24 hours (t-12, t-24, indicated in data as nlng t-12 a and nlng t-24 a) -and cells to which after withdrawal for 12 or 24 hours, subsequently GCSF was added back for 2 hours and 40 minutes (t-12+2h40m, t-24+2h40m, indicated in data as nlng t-12+2.4 a and nlng t-24+2.4 a). To obtain the data as presented in the manuscript the following was done:The values obtained for t0 were subtracted from t-12 and t-24. The values for t-12 and t-24 were subtracted from t-12+2h40m and t-24+th40m respectively. Subsequently the data was devided into genes that appeared to go down upon withdrawal of GCSF and up upon readdition and those with the converse gene expression pattern. Then a threshold of ?1.4 fold change was applied. If a certain gene obeyed these criteria for all described comparisons, the gene was considered a potential downstream target. The fold change in expression was calculated by 2^absolute value of subtracted normalised data.
Project description:This SuperSeries is composed of the following subset Series: GSE36816: GY118F downstream targets in iPS cells GSE36817: GY118F downstream effect in EpiSCs Refer to individual Series
Project description:This microarray was performed to gain insight in the effect of GY118F stimulation in EpiSCs. This array is part of the following paper to be published in Nature Communications: “JAK/STAT3 signalling is sufficient and dominant over antagonistic cues for the establishment of naïve pluripotency” by Anouk L. van Oosten, Yael Costa, Austin Smith & José C.R. Silva Gy0 a, gy0 b, gy0 c (triplicate samples for GY118F EpiSCs in N2B27+Activin+Fgf) c0 a, c0 b, c0 c (triplicate samples for Control EpiSCs in N2B27+Activin+Fgf) gy3 b, gy3 c (duplicate samples for GY118F EpiSCs in N2B27+Activin+Fgf stimulated with GCSF for 3 hours) c3 a, c3 b, c3 c (triplicate samples for Control EpiSCs in N2B27+Activin+Fgf stimulated with GCSF for 3 hours) Within the manucript genes of interest were defined as those that were significantly upregulated (P<0.01) above a 1.4 fold change threshold in GCSF stimulated GY118F EpiSCs as compared to those not stimulated, but for which the expression did not significantly change upon GCSF stimulation in the control. The fold change in expression was calculated by 2^absolute value of subtracted normalised data.
Project description:H9 human pluripotent stem cells with a stable Retinoblastoma-like protein 2 (Rbl2) knockdown or cells expressing a control Scramble shRNA were differentiated to neuroectoderm by inhibiting Activin/Nodal signalling and treating with Fgf2. Cells were collected at day 4 of differentiation and analysed for changes in global gene expression by microarray.
Project description:Identification of genes and pathways altered by PlaB, a bacterial natural product that acts as a spliceosome modulator by targeting the SF3b subunit of the spliceosome SKNMC, TC32, TC71, and RD-ES Ewing sarcoma cell lines were treated with 0.1% (v/v) DMSO vehicle or 5nM PlaB for 24 hours. Three samples in each group were analyzed.
Project description:The traditional method for studying cancer in vitro is to grow immortalized cancer cells in two-dimensional (2D) monolayers on plastic. However, many cellular features are impaired in these unnatural conditions and big alterations in gene expression in comparison to tumors have been reported. Three-dimensional (3D) cell culture models have become increasingly popular and are suggested to be better models than 2D monolayers due to improved cell-to-cell contacts and structures that resemble in vivo architecture. The aim of this study was to develop a simple high-throughput 3D drug screening method and to compare drug responses in JIMT1 breast cancer cells when grown in 2D, in polyHEMA coated anchorage independent 3D models and in Matrigel on-top 3D cell culture models. We screened 102 compounds with multiple concentrations and biological replicates for their effects on cell proliferation. The cells were either treated immediately upon plating or they were allowed to grow in 3D for four days prior to the drug treatment. Big variations in drug responses were observed between the models indicating that comparisons of culture model influenced drug sensitivities cannot be made based on effects of a single drug. However, we show with the 63 most prominent drugs that, in general, JIMT1 cells grown on Matrigel were significantly more sensitive to drugs than cells grown in 2D cultures, while responses of cells grown in polyHEMA resembled those of 2D. Furthermore, comparison of gene expression profiles of the cell culture models to xenograft tumors indicated that cells cultured in Matrigel and as xenografts most closely resembled each other. In this study we also suggest that 3D cultures can provide a platform for systematic experimentation of larger compound collections in a high-throughput mode and be used as alternatives for traditional 2D screens towards better comparability to in vivo state. Gene expression analysis of JIMT1 breast cancer cells cultured as xenografts for 43 days, in two dimensional cultures for seven days (2D7d), in polyHEMA three dimensional cell culture models for four and seven days (PH7d and PH7d), and in Matrigel three dimensional cultures for four and seven days (MG4d and MG7d). Two biological replicates was included for each sample.
Project description:The traditional method for studying cancer in vitro is to grow immortalized cancer cells in two-dimensional (2D) monolayers on plastic. However, many cellular features are impaired in these unnatural conditions and big alterations in gene expression in comparison to tumors have been reported. Three-dimensional (3D) cell culture models have become increasingly popular and are suggested to be better models than 2D monolayers due to improved cell-to-cell contacts and structures that resemble in vivo architecture. The aim of this study was to compare gene expression patterns of MCF7 breast cancer cells when grown as xenografts, in 2D, in polyHEMA coated anchorage independent 3D models and in Matrigel on-top 3D cell culture models. Surprisingly small variations in gene expression patterns were observed between the models indicating that 3D and xenograft are not always that different from 2D cell cultures. Gene expression analysis of MCF7 breast cancer cells cultured as xenografts for 43 days, in two dimensional cultures for seven days (2D7d), in polyHEMA three dimensional cell culture models for four and seven days (PH7d and PH7d), and in Matrigel three dimensional cultures for four and seven days (MG4d and MG7d). Two biological replicates was included for each sample.
Project description:A microarray study to compare gene expression during ovarian follicle development to better understand transcriptional changes during follicular growth. Multi-layered secondary follicles (150-180 M-BM-5m in diameter) were mechanically isolated from ovaries of 16-day old CD-1 mice and individually encapsulated in 0.5% alginate. Follicles were culture for up to 8 days. Day 0 samples correspond to follicles that were isolated and immediately frozen. Samples were collected after 0, 2, 4, 5, 6 and 8 days in culture in triplicate and day 0 was collected 5 times.
Project description:The objective of this study was to investigate the role of GATA6L with or without TNF stimulation. Microarrays were used to capture the transcriptional response to GATA6L overpexression at intermediate times after TNF stimulation. Cells were induced for GATA6L overexpression and evaluated at two hours after stimulation with saturating TNF in biological triplicate.