2.4 Total mRNA isolation and Dot-blot hybridization
The Ambion® Poly(A) Purist™ MAG kit (Invitrogen, USA) was taken for total mRNA isolation according to the manufacturer’s instructions. Dot blotting was performed as described previously (Zhang et al., 2021a). The aliquots (10 μL) of each RNA sample was spotted on a Hybond-N+ membrane (Amersham, USA), and cross-linked by UV (1200 × 100 μJ/cm2). Hybridization was performed for 1 h at 4 °C with anti-m6A antibody (#ab208577; Abcam) in 1×PBST buffer. The membrane was incubated with alkaline phosphatase-conjugated anti-mouse antibody (#ab98712; Abcam), the specific bands were visualised by adding nitro blue tetrazolium/5-bromo-4-chloro-3-indolyl phosphate as the substrates (Sangon Biotech, China).