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Fig. 3 | Virology Journal

Fig. 3

From: Host Cell Copper Transporters CTR1 and ATP7A are important for Influenza A virus replication

Fig. 3

Knockdown of host copper homeostasis genes affects viral macromolecular accumulation. (a) 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. (b) esiRNA treated cells were transfected with VPOL, firefly luciferase minigenome, and Renilla luciferase expression vectors. Relative viral RNA replication was assessed by normalizing firefly luciferase activity to Renilla luciferase activity. Biological replicates are displayed as points, with the mean and standard deviation of six biological replicates indicated with whiskers. (c and d) At 12 h.p.i. cells were lysed and proteins extracted. Proteins were subjected to western blotting using antisera to (c) viral NP protein, or (d) viral M1 protein. Relative viral proteins in knockdown immunoblots were quantified by densitometry. After transfer, gels were stained with Coomassie Brilliant Blue; a section is shown as loading control

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