RNA Gel Analysis of Small RNAs

RNA gel analysis of small RNAs was performed as described^{63 (link)}. Ten micrograms of total RNAs were separated on 15% polyacrylamide/8 M urea gels. After gel electrophoresis, the RNAs were transferred to a Hybond-NX nylon membrane (GE healthcare). Antisense complementary oligonucleotides (Supplementary Table 2) were synthesized with both 5’ and 3′ end-labeled biotin. A probe complementary to U6 (5′ CATCCTTGCGCAGGGGCCA 3′) was used to detect U6 as an internal control. Hybridization was performed for 16 h at 55 °C followed by washes. Signals were detected using the chemiluminescent nucleic acid detection module (Thermo Fisher, 89880).