Fig. 1

Addition of copper or chelator inhibits viral growth but not macromolecular accumulation. A549 cells’ growth medium was supplemented with 50 μM CuCl₂ or 10 μM TTM, to alter intracellular copper concentration, 24 hours before infection with influenza A/WSN/33 (H1N1). (a) At the indicated times post infection, infectious particles released into the medium were determined by immunostain assay. A representative data set is shown (i), with mean and standard deviation from the control condition for 3 independent experiments shown in accompanying panel (ii). Titer = fluorescent center forming units per mL. (b) At the indicated times cells were lysed and RNA extracted. Relative viral RNA amounts were determined by qRT-PCR with primers for the NP (segment 5) RNA and the host 18S rRNA. The means of two technical replicates are displayed as points, with the mean and standard deviation of three biological replicates indicated with whiskers. (c) At 12 h.p.i. cells were lysed and proteins extracted. Proteins were subjected to western blotting using antisera to the viral NP protein. A representative blot of at least 3 independent experiments is shown. After transfer the gels were stained with coomassie blue; a section is shown as loading control