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).