Identification of TDRKH-AS1 Interacting Proteins

TDRKH-AS1 and its antisense RNA were made from transcription with the Biotin RNA Labeling Mix (Roche, USA) and T7 RNA polymerase (Roche, USA). Biotinylated RNA was incubated with HUVECs nuclear extracts, and pulldown proteins were run on SDS-PAGE gels (Sigma, USA) and stained with silver staining solution (Beyotime, China) as our previous work [31 (link)]. The lncRNA pulled-down complex was sent to the company (Wininnovate Bio, China), using mass spectrometry to detect proteins interacting with TDRKH-AS1. We determined the proteins interacting with TDRKH-AS1, satisfying the criteria: only exists in the sense probe pulled-down complex and with at least 3 peptides. KEGG pathway enrichment analysis and GO (Gene Ontology) enrichment analysis were performed by clusterProfiler.