The FLI portion of EWS/FLI contributes a transcriptional regulatory function that is distinct from its DNA-binding function in Ewing sarcoma [CUT&RUN]

The purpose of this study was to compare the relevance of the FLI portion for genomic localization of the EWS/FLI protein in Ewing sarcoma cells. EWS/FLI cDNA constructs including various deletions of the FLI portion were virally transduced into cells and we sought to determine the ATAC profile of each. Endogenous EWS/FLI was knocked-down in the Ewing sarcoma A673 cell line and EWS/FLI cDNA constructs were expressed in these cells. Cells were collected after 8 days and antibody-directed micrococcal nuclease was used to determine the binding locations of these EWS/FLI protein constructs. Two biological replicates for each sample were used to construct cDNA libraries; each library was submitted twice to acheive sufficient read-depth and combined for analysis.