ABSTRACT: R-spondins (Rspos) comprise a family of four secreted proteins that have important roles in cell proliferation, cell fate determination and organogenesis. Rspos typically exert their effects by potentiating the Wnt/β-catenin signaling pathway. To systematically investigate the impact of Rspo/Wnt on gene expression, we performed a microarray analysis using C57MG mouse mammary epithelial cells treated with recombinant Rspo2 and/or Wnt3a. This study compares gene expression in C57MG cells after 24h treatment with recombinant Rspo2 and recombinant Wnt3a alone or in combination versus BSA treatment. Control cells were treated with BSA, since it was used as carrier for the recombinant proteins. Each treatment was done in biological triplicate.
Project description:Wnt signaling is upregulated frequently in several cancers, including sarcomas. Since, there is cell-context dependent variation in the target gene expression, to identify canonical Wnt targets in sarcomas, we used human mesenchymal stem cells. Human mesenchymal stem cells were treated with 100ng/ml recombinant Wnt3a either 6 hrs or 24 hrs. Total RNA was extracted from untreated and Wnt3a treated samples using Qiagen's RNA extraction kit.
Project description:Wnt signaling is upregulated frequently in several cancers, including sarcomas. Since, there is cell-context dependent variation in the target gene expression, to identify canonical Wnt targets in sarcomas, we used human mesenchymal stem cells. Overall design: Human mesenchymal stem cells were treated with 100ng/ml recombinant Wnt3a either 6 hrs or 24 hrs. Total RNA was extracted from untreated and Wnt3a treated samples using Qiagen's RNA extraction kit.
Project description:CXXC5 inhibits the canonical Wnt signaling pathway Impact of CXXC5 on Wnt stimulated gene expression in K562 cells Overall design: K562 cells were incubated with recombinant Wnt3a protein after overexpression (transient transfection of pEGFP-CXXC5 vector) or knockdown of CXXC5 (stable lentiviral infection of CXXC5 shRNA (pLKO.1; clone TRCN0000144558 and TRCN0000142729)
Project description:We analyzed the transcriptome of two different triple negative breast cancer (TNBC) cell lines to define a comprehensive list of Wnt target genes. Cells were treated with Wnt3a for 6h, 12h or 24h. We found up-regulated and down-regulated genes in response to Wnt3a treatment. They are involved in the Wnt pathway itself, and also in TGFß, p53 and Hedgehog pathways. Thorough characterization of these novel potential Wnt target genes may reveal new regulators of the canonical Wnt pathway. The comparison of our list of Wnt target genes with those published in other cellular contexts confirms the notion that Wnt target genes are tissue-, cell line- and treatment-specific. Cells were seeded in six-well plates, serum starved overnight then treated with Wnt3a for the indicated times (6, 12 and 24 hours). Triplicates for each condition were included in the experiment.
Project description:To test the potential of canonical Wnt signalling to modulate nociception via transcriptional regulation in dorsal root ganglion (DRG) neurons, we performed a genome-wide transcriptome analysis of neuron-enriched DRG cultures treated with Wnt3a or vehicle for 6 hours. Total RNA obtained from neuron-enriched mouse DRG culture subjected to Wnt3a treatment for 6 hours was compared to a matched control (vehicle-treated) DRG culture.
Project description:The observation that Tcf3 (MGI name: Tcf7l1) bound the same genes as core stem cell transcription factors, Oct4 (MGI name:Pou5f1), Sox2 and Nanog, revealed a potentially important aspect of the poorly understood mechanism whereby Wnts stimulate self renewal of pluripotent mouse embryonic stem (ES) cells. Although the conventional view of Tcf proteins as the β-catenin-binding effectors of Wnt signaling suggested Tcf3 should activate target genes in response to Wnts, here we show that Wnt3a and Tcf3 effectively antagonize each other’s effects on gene expression. Genetic ablation of Tcf3 caused similar effects as treating cells with recombinant Wnt3a. Moreover, Tcf3 was not necessary for Wnt3a-stimulation of gene expression as the majority of Wnt3a-stimulated genes exhibited a greater increase in Tcf3-/- ES cells than in Tcf3+/+ ES cells. These expression data, together with genetic experiments, show that Wnt3a stimulates ES cell self renewal by inhibiting Tcf3. Tcf3+/+ and Tcf3-/- mouse embryonic stem cells were cultured in self renewal conditions containing recombinant Wnt3a for RNA extraction and hybridization on Affymetrix microarrays.
Project description:In order to gain insight into the molecular events operating downstream of canonical wnt-signaling in myoblasts, we compared by microarray analysis the transcriptome of myoblast cultured for 4 hours in the presence and absence of Wnt3a. Overall design: Primary myoblasts isolated from newborn mice were treated for 4 hours in DMEM 2% horse serum in presence or in absence of Wnt3a (100ng/ml)
Project description:Wnt signaling is a major regulator of osteoblast differentiation and function. To investigate how Wnt3a signaling regulates osteoblastic gene expression and to identify the role of Lrp5 and Lrp6 in mediating Wnt3a signaling in osteoblasts, neonatal calvarial osteoblasts isolated from C57Bl6 (WT) and osteoblasts lacking Lrp5 (Lrp5KO), Lrp6 (Lrp6KO) and, both Lrp5 and 6 (Lrp5/6KO) were treated with Wnt3a for 24 hours and gene expression changes were quantified by RNA-seq. Overall design: Transcriptome analysis to identify Wnt3a regulated genes in Wild Type, Lrp5KO , Lrp6KO and Lrp5/6KO osteoblasts
Project description:The aim of the study was to characterize at a molecular level (changes in mRNA level) the effects of WNT3A on the human HepaRG hepatocellular carcinoma cell line. This was adressed by culturing HepaRG cells in presence or absence of Wnt3a. Overall design: HepaRG cells were treated with 200 ng/ml Wnt3a for 72hrs. To dissect the gene networks involved, experiments were also performed to silence β-catenin expression with a specific siRNA and to inhibit Wnt3a in the extracellular space with FZD8_CRD, a soluble Wnt-binding domain with SFRP-like functions. HepaRG cells were plated three days before siRNA transfection and treated with 200 ng/ml Wnt3a alone and/or 300 ng/ml FZD8_CRD from day one through day three after transfection. Independent culture experiments were performed in triplicate.
Project description:Wnt pathway is dysregulated in CLL-We characterized Wnt pathway gene expression in normal B and CLL-B cells and identified Wnt targets in normal B and CLL-B cells through this data set. In this dataset, we included normal B cells and CLL-B cells for Wnt pathway gene expression. This leads to the identification of 62 Wnt pathway components which are differnetially expressed between normal and CLl-B cells. We also included normal B cells and CLL-B cells with or without Wnt3a treatment and identified 468 and 676 Wnt regulated genes in normal and CLL B cells, respectively. 24 normal B cells and 179 CLL-B cells were used in the Wnt pathway gene expression analysis. Three normal and three CLL samples were included in the identification of Wnt regulated targets