Fig. 7

ELF3-AS1 directly interacts with the ILF2/ILF3 protein complex. A RNA pull-down assay for identification of ELF3-AS1 binding proteins was conducted. The sense (S) and anti-sense (AS) of ELF3-AS1 RNA were biotinylated, refolded, and incubated with SGC7901 cell lysates. B, C Identification of ELF3-AS1 binding proteins using a combined RNA pulldown-MS assay. According to the MS analysis of band labeled with a red arrow, RINI, TARBP2, ILF2 and ILF3 were the most possible RNA-binding proteins that interacted with ELF3-AS1. D Validation of ELF3-AS1 binding proteins obtained by RNA pulldown using western blotting. E Schematic workflow of the CHIRP assay for identification of ELF3-AS1 binding proteins. F The Venn plot of the ELF3-AS1 binding proteins identified by three mass spectrometry. G Verification of ILF2 and ILF3 protein with western blotting in the precipitation pulled down by different length of ELF3-AS1 RNA fragments. H ELF3 expression level in GC cells transfected with different lengths of ELF3-AS1 RNA fragments. I RIP-qPCR assay verified the interaction between ELF3-AS1 and ILF2/ILF3. Fold Enrichment was determined relative to IgG control. J RIP-seq_anti-ILF3 (GSE163815) assay verified the interaction between ILF3 protein and ELF3/ELF3-AS1 mRNA. **, P < 0.01