Figure 3.

Modulating Agrobacterium Infiltration Densities Impacts Both Accumulation Levels and Degree of Coexpression.

(A) Fluorescence images of leaves infiltrated with _pro35S:PHB-YFP and _pro35S:ZPR3-mCherry Agrobacterium strains at optical densities of 1 and 0.3, respectively, show relative PHB-YFP fluorescence is increased, but the frequency of cells accumulating both PHB-YFP and ZPR3-mCherry (gray hatched bar) is decreased, with more cells expressing exclusively PHB-YFP (green) or ZPR3-mCherry (magenta). Arrows, examples of bifluorescent cells; arrowheads, examples of unifluorescent cells.

(B) Representative TIRF images of PHB-YFP and ZPR3-mCherry SiMPull show that when PHB-YFP IP and co-IP spots are at single-spot resolution, ZPR3-mCherry IP spots are not.

(C) Histogram shows PHB-YFP IP spots are enriched over background, while the number of ZPR3-mCherry IP spots remains saturated (sat). Co-IP spot numbers for both ZPR3-mCherry and PHB-YFP are only marginally higher than background, suggestive of minimal coimmunoprecipitation. Green, PHB-YFP; magenta, ZPR3-mCherry. Spot densities (mean ± se) are calculated from at least three independent biological replicates.

Bars = 150 µm in (A) and 5 µm in (B).