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. 2013 Feb 19;8(2):e57063. doi: 10.1371/journal.pone.0057063

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© 2013 Chinnakannan et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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A549 cells were infected with RPV-Sa at a MOI of 5 or left uninfected. Eighteen hours post-infection cells were mock treated or treated with 1000 IU/ml of IFNα for 15 minutes, lysed in SDS-PAGE sample buffer containing protease and phosphatase inhibitors and the levels of phosphorylated Jak1 and Tyk2 were analyzed in Western blots using rabbit anti-Jak1P and rabbit anti-Tyk2P antibodies respectively. The blots were also probed with rabbit anti-Jak1, rabbit anti-Tyk2 and rabbit anti-RPV P protein (MB18) to analyze the total Jak1 and Tyk2 and to confirm RPV infection. The primary antibodies were detected with peroxidase-labelled anti-mouse or anti-rabbit IgG antibody. PCNA levels served as loading control.