circMAN1A2_009-pulled down proteins in HeLa cells

CircRNA pulldown assays were conducted to identify proteins interacting with circMAN1A2_009. In summary, HeLa cell protein extracts were incubated with biotin-labeled circMAN1A2_009 probes obtained from Tsingke Biotechnology, China, at room temperature for 1h. Subsequently, streptavidin magnetic beads were introduced into the cell lysates containing the probes and incubated overnight at 4°C. After three washes, proteins bound to the streptavidin magnetic beads were eluted using the Elution Buffer as per the instruction provided by the PierceTM Magnetic RNA-Protein Pull-Down Kit (Thermo, 20164, USA). The retrieved proteins underwent analysis through liquid chromatography-tandem mass spectrometry (LS-MS/MS) conducted by OE biotech Co. Ltd (Shanghai, China), or Western Blot.