Fig. 3
Accumulation of NS1 in virus-infected CLEC213 cells. Subconfluent CLEC213 cells in a 24-well plate were infected (0.5 PFU/cell) and lysed at the indicated times post-infection. Proteins in the lysates (0.8% of the 100 μl-lysate) were separated through SDS-PAGE, then transferred to a nylon membrane. For NS1 quantification, increasing amounts of recombinant NS1 protein (16 to 500 femtomoles) were loaded on the same gel 20 min after the lysates. Except for a C-terminal (His)6 extension, the recNS1 is identical to the viral NS1; its apparently slower migration profile only results from the delayed loading. NS1 was revealed through immunoblot, using a polyclonal rabbit serum. Lower panel: the same lysates were used for the western-blot detection of NP (the upper band corresponds to an unidentified cellular protein that was also detected in non-infected cells)