ewing sarcoma cell lines rd es  (DSMZ)

Journal: Life Science Alliance

Article Title: The chromatin landscape of primary synovial sarcoma organoids is linked to specific epigenetic mechanisms and dependencies

doi: 10.26508/lsa.202000808

Figure Lengend Snippet: (A) Gene dependency score analysis for a panel of PRC1 members in SyS versus all other cell lines present in the DepMap database identifies USP7 as a selective vulnerability in SyS. Genes shown in red had an adjusted P -value < 0.05. (B) Heat map showing ChIP-seq signals for USP7 at 4877 SS18-SSX–binding sites in C3H10T1/2 cells. 20-kb windows centered on SS18-SSX–binding sites are shown. (C) Representative example of USP7 recruitment by SS18-SSX at its binding sites in C3H10T1/2 cells. (D, E) Proximity ligation assay analysis demonstrates direct interaction between USP7 and SS18-SSX in C3H10T1/2 and HSSYII SyS cells. (F) Proximity ligation assay shows a significant decrease in interactions between the ncPRC1 members RING1B and RYBP following USP7 removal in HSSYII cells. (G) Cell viability assays in SyS (HSSYII, SYO1) and EwS (A673, RDES) cells upon CRISPR-mediated USP7 KO confirm the selective detrimental effect of USP7 depletion in the SyS cells. *** indicates P -value < 0.0001. Statistical analyses were performed by t test. See also .

Article Snippet: C3H10T1/2 cell line, Ewing sarcoma cell lines RDES and A673, MET5A and MRC5 cells were obtained from American Type Culture Collection and were cultured according to American Type Culture Collection recommendations.

Techniques: ChIP-sequencing, Binding Assay, Proximity Ligation Assay, CRISPR

Journal: Life Science Alliance

Article Title: The chromatin landscape of primary synovial sarcoma organoids is linked to specific epigenetic mechanisms and dependencies

doi: 10.26508/lsa.202000808

Figure Lengend Snippet: (A) Immunoblots showing that the expression of SS18-SSX-V5 does not change USP7 protein levels in C3H10T1/2 cells. Tubulin is used as a loading control. (B) Pie charts showing genomic distribution of USP7-binding sites in control and SS18-SSX–expressing C3H10T1/2 cells. (C) Venn diagrams showing the overlap between USP7-binding sites in control and SS18-SSX–expressing C3H10T1/2 cells. (D) Immunoblots confirm USP7 knockout in synovial sarcoma (HSSYII and SYO1) and Ewing sarcoma (A673 and RDES) cell lines. Actin is used as a loading control and densitometric analysis is reported as a percentage. (E) Effect of the USP7 inhibitor FT827 on growth and viability of Ewing sarcoma (A673) and synovial sarcoma (HSSYII and SYO1) cell lines, as well as synovial sarcoma organoids (SyS1). Cells were seeded in 100 mm plates at low concentration and treated for 72 h with the indicated amount of FT827 or vehicle (DMSO). The number of viable cells was evaluated by a trypan blue exclusion assay using an automated cell counter Countess II (Thermo Fisher Scientific). Mean values of triplicate counts are shown as a percentage with the mean value for vehicle treated cells set as 100%. Source data are available for this figure.

Article Snippet: C3H10T1/2 cell line, Ewing sarcoma cell lines RDES and A673, MET5A and MRC5 cells were obtained from American Type Culture Collection and were cultured according to American Type Culture Collection recommendations.

Techniques: Western Blot, Expressing, Control, Binding Assay, Knock-Out, Concentration Assay, Trypan Blue Exclusion Assay

Journal: Cell reports

Article Title: Systematic multi-omics cell line profiling uncovers principles of Ewing sarcoma fusion oncogene-mediated gene regulation

doi: 10.1016/j.celrep.2022.111761

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Ewing sarcoma cell line RD-ES , DSMZ, Braunschweig, Germany , RRID: CVCL_2169.

Techniques: Plasmid Preparation, Polymer, Virus, In Vivo, Recombinant, Reverse Transcription, DNA Methylation Assay, Knockdown, Gene Expression, Microarray, shRNA, Software

Journal: Artificial cells, nanomedicine, and biotechnology

Article Title: Integrated bioinformatics analysis of miRNA expression in Ewing sarcoma and potential regulatory effects of miR-21 via targeting ALCAM/CD166.

doi: 10.1080/21691401.2019.1620760

Figure Lengend Snippet: Figure 11. (A) Schematic diagram of luciferase reporter assay. The green dots represent stronger fluorescence signal, the blue dots represent weak fluorescence sig- nal. (B) Relative luciferase activity of indicated ALCAM reporter constructs in 293 T cells. Transfection of miR-21 suppressed the activity of the luciferase reporter which contained the 30-UTR of ALCAM significantly but did not influence the luciferase activity of the mutated 30-UTR structure in 293 T cells. p < .01. (C) Microscopic observation of hMSC cell line, A673 cell line and 293 T cells.

Article Snippet: Cell lines and cell culture The human Ewing sarcoma cell lines A673 SK-ES-1 RD-ES and the hMSC cell line was obtained from the American Type Culture Collection (ATCC).

Techniques: Luciferase, Reporter Assay, Fluorescence, Activity Assay, Construct, Transfection