Project description:To analyse whether the activation of TGF-ß signalling (recombinant hTGF-ß, 5 ng/mL; PeproTech) is able to invoke a full phenotype- switch, we performed RNA sequencing of melanoma cultures from two independent melanoma cell lines at two different time point after TGF-β1 (5 ng/ml, preprotech).
Project description:The current study examines the functional role of VEPH1, a PH domain containing protein and the human ortholog of Drosophila melted, in ovarian cancer cell lines. Elevated VEPH1 is associated with FOXO and Hippo signaling and was found to suppress TGF-ß induced target genes. SKOV3 cell lines were established as either CdCl2 inducible Flag-tagged VEPH1 expression or mock-transfected. Gene expression profiles were then generated both with and without the presence of TGF-ß to induce TGF-ß signaling. 3 replicates per treatment group were used.
Project description:By performing oligonucleotide microarray analysis the role of Smad4 in response to TGF-ß was evaluated in established MDA-MB-468 Smad4 negative and positive clones that were treated with TGF-ß for different time points. Keywords: time-course
Project description:To investigate the function of long noncoding RNA lncMGC in WT and lncMGC KO mouse kidney mesangial cells treated with TGF-ß. Assay for Transposase-Accessible Chromatin (ATAC) sequencing on 4 types of kidney messangial cell samples: isolated from wild type (WT) or lncMGC knockout (lncMGC_KO) mouses, each treated with either SD or TGF-ß, namely WT_SD, WT_TGF, KO_SD and KO_TGF.
Project description:To investigate the function of long noncoding RNA lncMGC in WT and lncMGC KO mouse kidney mesangial cells treated with TGF-ß. Chromatin immunoprecipitation DNA-sequencing (ChIP-seq) for histone modication mark H3K27ac on 4 types of kidney messangial cell samples: isolated from wild type (WT) or lncMGC knockout (lncMGC_KO) mouses, each treated with either SD or TGF-ß, namely WT_SD, WT_TGF, KO_SD and KO_TGF.
Project description:To investigate the function of long noncoding RNA lncMGC in WT and lncMGC KO mouse kidney mesangial cells treated with TGF-ß. We then performed gene expression profiling analysis using data obtained from RNA-seq of 4 types of kidney messangial cell samples: wild type (WT) or lncMGC knockout (lncMGC_KO) mouse, each treated with either SD or TGF-ß, namely WT_SD, WT_TGF, KO_SD and KO_TGF.
Project description:Myocardin-related transcription factors (MRTFs) are robust co-activator of Serum Reponse Factor, and regulate cell adhesion and motility by controlling the transcription level of cytoskeletal components. In tumor progression, MRTFs has been reported to contribute to metastatic potential. Previously, we reported thymosin-ß4 (Tß4, which is encoded by Tmsb4x gene) as an activator of MRTFs. Enhanced expression of Tß4 is frequently observed during tumor progression, and associated with poor prognosis. However, the relationships between MRTFs and Tß4 in tumor progression are largely unknown. Here we analyze the effect of Tß4 knockout on the transcription levels of MRTFs-target genes. In particular, we focused on their functions in TGF-ß signaling because both MRTFs and Tß4 are down-stream effectors of TGF-ß.
Project description:Argonaute (Ago) proteins, which act in post-transcriptional gene regulation directed by small RNAs, are vital for normal stem cell biology. Here we report the genomic characterization of stable Ago-deficient mouse embryonic stem cells (mESC) and determine the direct and system level response to loss of Ago-mediated regulation. We find mESCs lacking all four Ago proteins are viable, do not repress microRNA (miRNA)-targeted cellular RNAs, and show distinctive gene network signatures. Profiling of RNA expression and epigenetic activity in an Ago mutant genetic series indicates that early responses to Ago loss are driven by transcriptional regulatory networks, in particular the Tgf-ß/Smad transcriptional network. This finding is confirmed using a time course analysis of Ago depletion and Ago rescue experiments. Detailed analysis places Tgf-ß/Smad activation upstream of cell cycle regulator activation, such as Cdkn1a, and repression of the c-Myc transcriptional network. The Tgf-ß/Smad pathway is directly controlled by multiple low-affinity miRNA interactions with Tgf-ß/Activin receptor mRNAs and receptor-mediated activation is required for Tgf-ß/Smad target induction with Ago loss. Our characterization reveals the interplay of post-transcriptional regulatory pathways with transcriptional networks in maintaining cell state and likely coordinating cell state transitions.