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. 2011 Apr 1;6(4):e17796. doi: 10.1371/journal.pone.0017796

Figure 4. Co-immunoprecipitation (co-IP) of transiently expressed viral proteins.

Figure 4

HeLa cells were co-transfected with a combination of two plasmids expressing HA- and myc-tagged proteins. Panels (A) and (B) are inputs that show the expression of HA- and myc-tagged proteins detected by Western blot analysis with anti-HA and anti-myc antibodies, respectively. Lanes 1 and 9 are from the same cell lysate, likewise, lanes 2 and 10, and so on. Lanes 7, 8, 15, and 16 are negative controls based on our YTH data that revealed no interactions. Panels (C) and (D) represent co-IP experiments (IP-anti-HA) with anti-HA agarose antibodies and visualized by Western blot analysis using anti-HA and anti-myc antibodies, respectively. Panels (E) and (F) are co-IP experiments (IP-anti-myc) with anti-myc agarose antibodies (IP-anti-Myc) and visualized by Western blot analysis using anti-HA and anti-myc antibodies, respectively. The protein species, which migrated at approximately 82 kDa and was detected by the anti-HA antibody in the input protein samples of the cell lysate (A) but not in the protein samples immunoprecipitated with either the anti-HA or anti-myc antibodies (C and E), may represent a cellular protein that non-specifically reacts with the anti-HA antibody.