Northern blot analyses
A 30–45 µg aliquot of high-quality total RNA (A260/280=2.00) was
separated on a 19% polyacrylamide denaturing gel, and then transferred
to a Hybond membrane (GE healthcare) for 2 h at 200 mA at 4 °C. In
brief, after crosslinking for 5 min with ultraviolet irradiation, the
membrane was hybridized overnight at 45 °C with DNA probes (Yu et al.,
2011). Washing and autoradiography of the Hybond membrane were performed
according to the instructions of the North2South™ Chemiluminescent
Hybridization and Detection kit (No.17097, Pierce, Rockford, IL).