Skip to main content
. 2006 Oct 4;80(24):11968–11981. doi: 10.1128/JVI.01394-06

FIG. 7.

FIG. 7.

Enhancement of ORF56 expression by ORF57. (A) Detection of ORF56 transcripts by Northern blotting. Fractionated cytoplasmic (C) or nuclear (N) total RNA from 293 cells transfected with 1 μg of pVM9 (ORF56-FLAG) in the presence or absence of 0.2 μg of pVM7 (ORF57-FLAG) or pFLAG-CMV-5.1 control vector was prepared 24 h after transfection. Five micrograms of total RNA was used for Northern blot analysis with a 32P-labeled ORF56-specific probe. The same membrane was reprobed separately with a 32P-labeled GAPDH-specific probe and a U6-specific probe for sample loading and fractionation efficiency. The nuclear and cytoplasmic fractionation efficiencies were also verified by the presence or absence of 45S and 32S pre-rRNA. T, total RNA from unfractionated 293 cells. (B) Detection of ORF56-FLAG fusion protein by Western blot analysis. Protein samples were prepared after 24 h of transfection and blotted with anti-FLAG antibody to detect ORF56- and ORF57-FLAG fusions. The membrane was reprobed with anti-β-tubulin antibody for sample loading.