Project description:FECR1 is an exonic circular RNA of the proto-oncogene FLI1. FECR1 is identified as a molecular component in the FLI1 promoter chromatin complex. To study its function in breast cancer, we utilized a RNA reverse transcription-associated trap sequencing (RAT-Seq) approach to map the DNA binding targets in MDA-231 cells.
Project description:MDA-MB231 cells were transfected with control vector, RAI2-overexpression vector, or vector with RAI2-4A, which has a mutated interaction domain w.r.t. CtBP/CtBP2
Project description:MDA-MB231 cells were transfected with control vector, RAI2-overexpression vector, or vector with RAI2-4A, which has a mutated interaction domain w.r.t. CtBP/CtBP2 Three replicates each of control condition, RAI2 overexpression or RAI2-4A overexpression.
Project description:To determine the absolute copy number of proteins in MDA-MB231 breast cancer cells, we employed IBAQ mediated absolute quantification of proteins based on (Schwanhäusser et al., Nature, 2011), with some modifications. Maqquant calculated iBAQ values were calibrated using spike-in standards, and used to calculate copy numbers for each identified protein within the dataset. Copy numbers for a total of 3,584 proteins were calculated in MDA-MB231 cells.
Project description:G9a is an H3K9m2 methyltransferase, which is critical in controlling gene suppression and DNA methylation. We used microarray analysis to identify the target genes that are regulated by G9a in MDA-MB231 cells, in which E-cadherin is silenced. G9a expression was stably knocked-down in MDA-MB231 cells. RNA from this clone and parental (control) cells were purified for microarray analysis.
Project description:To define and compare the interactomes of the RNA binding protein HNRNPC in poorly vs. efficiently metastatic breast adenocarcinoma cells, we carried out immunoprecipitation of endogenous HNRNPC from parental MDA-MB231 cells vs. its highly metastatic isogenic derivate, the MDA-MB231-LM2 cells. We used a non-specific MOUSE IgG IP from each line as control. Each IP was performed in triplicate, and analysed by LC-MS/MS, on a Thermo Q-Exactive-plus instrument.
Project description:To test specificity of human microarray probes against mouse mRNA serially mixed human MDA-MB231 cancer cells' mRNAs with mouse Astrocytes were hybridized against human microarray and vice versa by changing the ratios of human and mouse samples from 100% to 0%. Mixed human: 1 MDA-MB231, 1 MDA231_Astro(91), 2 MDA231_Astro(73), 2 MDA231_Astro(55),2 MDA231_Astro(37),1 MDA231_Astro(19), 1 Astrocytes Mixed mouse: 1 MDA-MB231, 1 MDA231_Astro(91), 2 MDA231_Astro(73), 2 MDA231_Astro(55),2 MDA231_Astro(37),1 MDA231_Astro(19), 1 Astrocytes
Project description:UGT8 is the first key enzyme that catalyzes the transfer of galactose to ceramide for the synthesis of galactosylceramide. We used microarray analysis to identify the genes that are regulated by UGT8 in MDA-MB231 cells.
Project description:We analysed the impact of LARP6 depletion on the proteome of actively growing MDA-MB231 breast cancer cells by SILAC. For this purpose, Light (L) SILAC-labelled MDA-MB231 cells were treated with non-targeting control (NT) or two independent LARP6 siRNA (18i & 97i) for 72 hrs, before lysis in 4% SDS, 100mM Tris/HCl pH 7.5. In parallel, Heavy (H)SILAC labelled non-transfected MDA-MB231 cells were grown and lysed similarly. Each L labeled lysate was then mixed with an equal amount of H labelled lysate. Mixing of samples to the same H standard therefore allowed cross-comparison of different siRNA treatments from separate runs.
Project description:G9a is an H3K9m2 methyltransferase, which is critical in controlling gene suppression and DNA methylation. We used microarray analysis to identify the target genes that are regulated by G9a in MDA-MB231 cells, in which E-cadherin is silenced.