Fig. 2.

VHL interacts with SARS-CoV nsp16. (A) 6xHis-VHL or 6xHis-nsp16 proteins expressed in BL21 cells were incubated with glutathione beads and the purified GST, GST-nsp16 or GST-VHL proteins. The bound proteins were eluted and detected by Western blotting with an anti-His or anti-GST antibody. (B) Flag-nsp16 and HA-VHL were transfected into HEK293T cells. After 36 h, the cell extracts were immunoprecipitated (IP) with an anti-HA or anti-Flag antibody and analyzed by immunoblotting (IB) with an anti-Flag or anti-HA antibody (top panel). (C) The eGFP-nsp16 and Red-VHL plasmids were co-transfected into HeLa cells. After 24 h, the cells were fixed with 4% paraformaldehyde and the nuclei were stained with DAPI. The cells were examined by confocal microscopy. (D) After transfection, the Vero cells were fixed with 4% paraformaldehyde, labeled with rabbit anti-Flag antibody (1:100), then probed with FITC-tagged goat anti-rabbit IgG. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)