Circular RNA Interaction Profiling

The Magna RIP kit (Millipore, Bedford, MA, USA) was adopted for RIP assay according to the manufacturer’s instructions. Saos2 and MG63 cells were lysed by RIP lysis buffer (Beyotime, Shanghai, China) as previously described [30 (link)]. The whole cell extract (100 μL) and radio-immunoprecipitation assay (RIPA) buffer (Beyotime, Shanghai, China) were incubated together. The magnetic beads pre-coated with anti-argonaute 2 protein (Ago2) antibody (Abcam, Shanghai, China) were added to interact with circ_0078767-miR-889 complex, and anti-immunoglobulin G (IgG) antibody (Millipore, Bedford, MA, USA) served as the control. After the immunoprecipitate and proteinase K were incubated to remove the protein, qRT-PCR was utilized to determine the relative enrichment of circ_0078767 in the immunoprecipitate.