Impact of EWS knockdown on LTBR mediated Gene Activation in human fibroblasts

LTβR signaling is crucial for immune development, homeostasis, and inflammation. To identify protein determinants that control the quality of LTβR function, we use a novel proteomics approach to identify EWS as a novel signaling component of this pathway. LTβR signaling protein, TRAF3, controls the protein binding activity of EWS via formation of mutually exclusive protein-protein interactions. With transcriptomics and knockdown experiments we identify several pro-inflammatory genes with induction kinetics that are controlled by EWS during LTβR signaling. Given the link between gene induction responsiveness and mRNA decay, we show EWS controls the decay of these transcripts. Fibroblasts expressing control or EWS targeting shRNAs were stimulated with agonistic goat anti-human LTBR antibody for 0, 2, or 6 hours. Total RNA harvested and assayed using nCounter Human Inflammation panel V2