Drosha antibody

CHIR99021 and rabbit anti-β-catenin antibody were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). BIO was purchased from Sigma-Aldrich. β-Actin mouse monoclonal antibody was purchased from Beijing TransGen Biotech Co., Ltd (Beijing, China). Drosha antibody was purchased from Cell signaling (Danvers, MA). Alexa Fluor 555-labeled goat anti-rabbit IgG and anti-rabbit/mouse horseradish peroxidase-conjugated secondary antibody were obtained from the Beyotime Institute of Biotechnology (Jiangsu, China).

pcDNA3.1-MS2, pcDNA3.1-MS2-SPINT1-AS1, or pcDNA3.1-MS2-SPINT1-AS1-del was co-transfected with pMS2-GFP (Addgene) into HeLa cells. After culture for another 48 h, cells were used to conduct RIP assays using the Magna RIP RNA-Binding Protein Immunoprecipitation Kit (Millipore, Billerica, MA, United States) and a GFP antibody (5 μg per reaction; 11814460001, Roche). For anti-DROSHA, anti-DGCR8 RIP, HeLa cells with SPINT1-AS1 or SPINT-AS1-del stable overexpression and HeLa cells with SPINT1-AS1 stable silencing were used to conduct RIP assays using a DROSHA antibody (5 μg per reaction; #3410, Cell Signaling Technology, Danvers, MA, United States), a DGCR8 antibody (5 μg per reaction; ab191875, Abcam, Hong Kong, China), and the Magna RIP RNA-Binding Protein Immunoprecipitation Kit.