ABSTRACT: Response of HEK293-cells after transfection with EWS-FLI1. HEK293 cells were transfected with the expression vector pIRES2-EGFP containing type I EWS-FLI1 or empty control vector. For transient transfection standard DEAE dextran method was used and RNA was isolated 48h post transfection. For stable transfection cells were transfected using FuGENE 6 (Roche, Mannheim, Germany) and cells were selected with 400 ug/mL G418. DNA-microarray analysis was performed using Affymetrix HG-U133A microarrays.(see Staege et al. Cancer Res. 2004) Keywords = HEK293 Keywords = EWS-FLI1 Keywords = Ewing family tumors Keywords: other
Project description:Response of HEK293-cells after transfection with EWS-FLI1. HEK293 cells were transfected with the expression vector pIRES2-EGFP containing type I EWS-FLI1 or empty control vector. For transient transfection standard DEAE dextran method was used and RNA was isolated 48h post transfection. For stable transfection cells were transfected using FuGENE 6 (Roche, Mannheim, Germany) and cells were selected with 400 ug/mL G418. DNA-microarray analysis was performed using Affymetrix HG-U133A microarrays.(see Staege et al. Cancer Res. 2004)
Project description:Transient transfection of a Ewing's Sarcoma cell line expressing type I EWS-FLI1 fusion and doxycycline-inducible short hairpin RNA against EWS-FLI1 (A673sh) In total, 7 samples were analysed: empty vector control and two nuclear directed AKT- and CDK2- phosphorylation resistant FOXO1 versions as well as sh-scrambled and sh-FOXO1, either in the presence (w.o. Doxy.) or absence of EWS-FLI1 (+ Doxy.) each 2 replicates
Project description:Transient transfection of a Ewing's Sarcoma cell line expressing type I EWS-FLI1 fusion and doxycycline-inducible short hairpin RNA against EWS-FLI1 (A673sh)
Project description:Ewing’s sarcoma (EWS) is a cancer of the bones or soft tissues in children and adolescents. EWS-FLI1 is a transcriptional factor and the key driver of EWS. To characterize the changes of downstream transcriptional profiles of EWS-FLI1 in A673 cells upon knockdown of EWS-FLI1 and deubiquitinase USP9X, RNA-seq was performed in A673 cells transfected with EWS-FLI1 esiRNA, USP9X esiRNA or GFP esiRNA as control. The data indicates that USP9X regulates transcriptional profiles of EWS through mediating EWS-FLI1.
Project description:Response of A673-cells to transfection with interleukin 2 (IL-2). Ewing family tumor (EFT) cell line A673 was transfected with an expression vector for IL-2 (cell line SBVGA1) or empty control vector (cell line SBVGA3). Cells were harvested at 3 different time points and processed for DNA-microarray analysis using Affymetrix HG-U95A microarrays. Primary image analysis was performed using MAS 5.0. In addition, primary analysis was performed with MAS 4.0 and data were scaled to an target intesity of 200. These data have been analysed together with other published data that have been scaled to the same target intensity (see Staege et al. Pediatr Blood Cancer 2004 Jul;43(1):23-34. See also Staege et al. Cancer Res. 2004).
Project description:Expression profiles were generated from hESC-derived neural crest stem cells following transduction with GFP control vector or EWS-FLI1 vector. Expression was analyzed in stem cell conditions 5 days after transduction (undifferentiated conditions) and after 6 weeks in differentiation media (differentiation conditions). Changes in gene expression over time were compared between control and EWS-FLI1 expresssing cells. Total RNA was extracted from 3 replicates for each of 4 conditions (undifferentiated control, undifferentiated EWS-FLI1, differentiated control and differentiated EWS-FLI1. Samples were analyzed by Affymetrix exon arrays using standard procedures.
Project description:STAG2, a member of cohesin, is one of the most recurrently mutated genes in human cancer. Here, we investigated STAG2 function in the context of Ewing sarcoma, an aggressive bone tumor driven by EWS-FLI1 oncogene chimeric transcription factor. Five different Ewing sarcoma cell lines were transfected with siCT or 2 different siRNAs targeting STAG2 (siSA2#6 or siSA2#8). RNA in A673 & TC71 (siSTAG2) was extracted 24, 48 and 72 hours post-transfection. RNA in EW1, CHLA-10 and CHLA-258 (siSTAG2 and siEWSR1-FLI1) was extracted 72 hours post-transfection. RNA-seq based anlayses of these experiments highligted that STAG2 knock-down alters EWSR1-FLI1 activation signature.
Project description:We obtained transcriptome profiling (RNA-seq) of human esophageal squamous cell carcinoma cell line KYSE150 stabley transfected clones with pIRES2-EGFP vector or human NCCRP1-expression vector by using next generation sequencing.
Project description:We obtained transcriptome profiling (RNA-seq) of human esophageal squamous cell carcinoma cell line KYSE30 cells stabley transfected with pIRES2-EGFP vector or human RHCG-expression vector by using next generation sequencing.