Research Paper Volume 12, Issue 7 pp 5751—5763

Molecular anatomy of the subcellular localization and nuclear import mechanism of herpes simplex virus 1 UL6

Figure 5. Subcellular distribution of UL6 in presence of different shRNA expression plasmids. (A) Verification of knock down efficiency of the constructed shRNA expression plasmids for importin α1, importin α7 and transportin-1. HEK293T cells were co-transfected with the plasmids combination of Flag-kα2 (importin α1)/pSuper, Flag-kα2/shRandom, Flag-kα2/shImportin-α1, Flag-kα6 (importin α7)/pSuper, Flag-kα6/shRandom, Flag-kα6/shImportin-α7, pFLAG-CMV-transportin-1/pSuper, pFLAG-CMV-transportin-1/shRandom or pFLAG-CMV-transportin-1/shTransportin-1 for 24 h. Then, cells were lysed and IB was performed with anti-Flag mAb. β-actin was used as a loading control. (B) One or two or three plasmids of shImportin-α1, shImportin-α7 and shTransportin-1 were co-transfected with pFLAG-UL6 into COS-7 cells for 24 h, then IFA was carried out using confocal fluorescence microscopy. Statistical analysis of the fluorescence was shown in Table 4.