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. 2011 Feb 22;30(7):1343–1356. doi: 10.1038/emboj.2011.39

Figure 3.

Figure 3

Inactive TOR kinase mutant fails to stimulate TAV-mediated transactivation. (A) Schematic diagram of the monocistronic and dicistronic reporter constructs. (B) Arabidopsis mesophyll protoplasts were cotransfected with the two reporter plasmids pmonoGFP and pbiGUS, as well as effector plasmids expressing TAV and either TOR or its inactive derivative (D2247A) as indicated. Activity of GUS synthesized in protoplasts transfected with pTAV only was set as 100% (150 000 r.f.u.), and quantified GFP expression in protoplasts transfected with only reporter constructs—as 100%. biGUS mRNA levels were analysed by qPCR (grey bars; RNA value in the presence of pTOR(D2247A) was set at 100%), and semiquantitative RT–PCR 24 h after protoplast transformation. The data shown are the means of three independent assays: error bars indicate s.d. (C) TAV, TOR, S6K1 and S6K1-T449-P accumulation was analysed by western blot using appropriate antibodies.