Fig. 7.

Degradation of CAT3 protein in leaves agroinfiltrated with CAT3-FLAG and CMV 2b (Y2b or N2b). a Western blot analysis of the protein extracts from agroinfiltrated leaves at 3 dpi. CAT3-FLAG was detected using anti-CAT3 antibodies. Protein samples were prepared as essentially described in Masuta et al. (1995). Briefly, leaf tissue was homogenized in 1 ml of PBS/Tween and centrifuged at 10000×g for 20 min to separate the supernatant (S) and pellet (P). After Laemmli’s sample buffer was added to each of S and P, the samples were subjected to SDS-PAGE. Arrow indicates both the endogenous tobacco catalases and CAT3-FLAG proteins, and asterisk shows a possible breakdown product derived from the catalases. The blot was stained with amido black (AB) to confirm the equivalence of protein loading. b Western blot with anti-2b antibodies. Y2b and N2b were detected on the same blot. Arrows indicate the 2b protein. The loading controls of S and P lanes of Y2b and N2b are equal to lanes 5 and 6 (for Y2b) and lanes 7 and 8 (for N2b) of AB staining in a, respectively