EZ Magna RIP Kit (Millipore) was used for RNA immunoprecipitation (RIP) following the manufacturer’s instructions. Briefly, cells were lysed in complete RIP lysis buffer. Then the cell lysates were incubated with magnetic beads conjugated with an anti‑Ago2 antibody (1:2000, Abcam, USA) or a normal IgG antibody (1:2000, Abcam, USA) for 24 h at 4 °C. After that, the magnetic beads were washed and treated with proteinase K (50 μg/mL) at 37 °C for 1 h to remove protein. Finally, circSCAP, miR-7, and SMAD2 in immunoprecipitated RNAs were quantified by qRT‐PCR analysis (Qin et al. 2021 (link)).
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