2.14. RNA pull-down assay

In brief, 0.5 ml buffer (25 mM Tris-HCl, 0.05% NP-40, 70 mM KCl, 2.5 mM EDTA, 80 U/ml RNase inhibitor and 1× protease inhibitor) were used to incubate CHON-001 and ATDC5 on ice. After centrifugation for 20 min, the supernatant was immediately collected. Following that, biotin-coupled miR-138-5p (bio-miR-138-5p) or biotin-coupled control miRNA (bio-NC) (RiboBio, Guangzhou, China) was accordingly mixed with the supernatant and incubated for 2 h. The streptavidin magnetic beads were then loaded. 4 h later, the streptavidin magnetic beads were accordingly isolated, and the enrichment of SIRT1 mRNA enrichment was quantified via qRT-PCR in the isolated RNA.