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. 2013 May 9;8(5):e64202. doi: 10.1371/journal.pone.0064202

Figure 7. PIV5-V blocks the interaction between mda-5 and LGP2 to inhibit IFN induction.

Figure 7

(A) Yeast cells were transformed with a plasmid expressing the helicase domain of mda-5 as a GAL4DBD fusion, a plasmid expressing LGP2 as a GAL4AD fusion and either the empty vector pHON7 (−) or pHON7 expressing the V protein from PIV5 (PIV5-V). Positive transformants were selected on SD-L-W-U and subsequently streaked onto SD-L-W-U-H + 5 mM 3-AT to assay for an interaction between the GAL4DBD fusion protein and the GAL4AD fusion protein. (B) HEK293 cells were transfected with the IFN-β reporter plasmid, the β-galactosidase expression plasmid and either the empty vector pEFpl2 (−), pEF.LGP2 or pEF.PIV5-V as indicated. 24 hours after transfection cells were further transfected with 2 ng poly(I:C) for 16 hours. Cell lysates were analysed for luciferase and β-galactosidase activity, and relative expression levels calculated.