RNA Immunoprecipitation of MALAT1-miR-382-3p-BDNF-AGO2 Complex

The RIP kit (RIP-12RXN, Sigma-Aldrich, USA) was adopted to detect the binding between MALAT1, miR-382-3p, BDNF, and AGO2 protein. Upon reaching 80–90% confluency, the culture medium was removed. Then, the cells were lyzed using an equal volume of RIPA lysis buffer (P0013B, Beyotime) for 5 min and centrifuged at 14,000 rpm for 10 min at 4 ℃, with the supernatant collected. A portion of the cell extract was used as input, and the rest was incubated with antibodies for co-precipitation (Zhang et al. 2020 (link)). The RIP antibody we used was AGO2 (1:100, ab186733, Abcam, UK) and IgG (1:100, ab200699, Abcam, UK, negative control).