Protocol for in vitro phase separation of ARID3B proteins.

Liquid-liquid phase separation (LLPS) of ARID3B is critical in nonsyndromic cleft lip/palate pathogenesis. Here, we present a protocol to visualize ARID3B LLPS. We describe steps for regenerating LLPS with purified protein in vitro and detecting ARID3B condensates in mammalian cells. This protocol enables analysis of ARID3B phase separation dynamics and the study of other transcription factors that undergo LLPS. For complete details on the use and execution of this protocol, please refer to Li et al.1.