Cellular and Zebrafish RNA Extraction

For cellular RNA extraction, 400 µL TRIzol was added to the cell culture wells for lysis. For zebrafish RNA extraction, 10 zebrafish were placed into 1.5-mL EP tubes, and 400 µL TRIzol added for lysis (38 (link)). RNA was isolated and purified according to the manufacturer’s instructions for the use of TRIzol reagent (TaKaRa, Japan). RNA was reverse transcribed into cDNA using a cDNA Reverse Transcription Kit (Monad, Shanghai, China). Real-time quantitative PCR (RT-qPCR) analysis was performed using SYBR Green PCR Mix (Monad, Shanghai, China) in the ABI QuantStudio 3 PCR system (Applied Biosystems, Waltham, MA, USA). The primers used are listed in Table 1.