Ebola virus VP35 has novel NTPase and helicase-like activities

Shu, Ting; Gan, Tianyu; Bai, Peng; Wang, Xiaotong; Qian, Qi; Zhou, Hui; Cheng, Qi; Qiu, Yang; Yin, Lei; Zhong, Jin; Zhou, Xi

Northern blot was performed as previously described (42) with minor modification. Five g of total RNAs were electrophoresed on denaturing 1.2% agarose gels with 2.2 M formaldehyde, and then capillary transferred to Hybond-A nylon membrane (GE Healthcare). The membranes were hybridized with the DIG-labeled RNA probe at 65 • C for 12 h, and then incubated with anti-DIG-alkaline phosphatase antibody (Roche), followed by incubation with CDP-STAR (Roche) at 37 • C for 10 min. The signals were detected by radiography on X-ray film (FujiFilm, Tokyo, Japan). The DIG-RNA probe targeting the negative-strand of EBOV 3 UTR 1-200 nt were produced via in vitro transcription using DIG RNA labeling mix (Roche).