Figure 8B Co-IP analyses to analyze interactions of NbREM1 with γb or γb3CS in N. benthamiana leaf tissues infiltrated with A. tumefaciens mixtures containing movement-deficient BSMV plasmids (RNAα + RNAγγb-GFP or RNAγ3CS-GFP) and NbREM1-3xFlag. Total proteins were extracted at 3 dpi and immunoprecipitated with anti-Flag beads. Input and immunoprecipitated proteins (IP) were analyzed by Western blot analyses with an anti-GFP or anti-Flag antibodies. Data information: In B, , representative data are shown and three biological replicates had similar results.D Analysis of the cleaved 5(6)-carboxyfluorescein (CF) movement in N. benthamiana leaves. RNAα + RNAγ/RNAγ3CS were co-infiltrated into transiently overexpressed NbREM1 N. benthamiana leaves, and 1 μL CFDA was loaded onto the adaxial leaf blade surface and incubated for 5 min followed by confocal analysis at 3 dpi. The areas of dye diffusion are marked by dashed white circles. Scale bars, 100 μm. Data information: In D, representative data are shown and three biological replicates had similar results.E Quantification of CF movement. Dye diffusion areas identified by the dashed white circles in panel D were measured by image J software (n = 6). Data information: In (E, the box boundaries indicate the upper (25th percentile) and lower (75th percentile) quartiles, whiskers indicate minimum and maximum values, central band indicates the median. Letters in the chart denote statistically significant differences among groups according to the Duncan's multiple range test (p < 0.05).F Aniline blue staining of callose in N. benthamiana leaf tissues. Leaves were infiltrated with mixtures of different A. tumefaciens constructs indicated beside the panels. At 3 dpi, the 0.1% aniline blue fluorochrome was infiltrated into N. benthamiana leaves. After incubation at 5 min in the dark, the leaves were observed by confocal microscopy at 405 nm. Scale bars, 20 μm. Data information: representative data are shown and three biological replicates had similar results.G Quantification of callose staining intensity as shown in panel F. The relative callose staining intensity was set to 1 in the REM samples (n = 10). Data information: In , the box boundaries indicate the upper (25th percentile) and lower (75th percentile) quartiles, whiskers indicate minimum and maximum values, central band indicates the median. Letters in the chart denote statistically significant differences among groups according to the Duncan's multiple range test (p < 0.05).H Aniline blue staining of callose in leaves with different treatments above the panel in NbPAT15 and NbPAT21 RNAi plants. Scale bars, 20 μm. Data information: In H), representative data are shown and three biological replicates had similar results.I Quantification of callose staining intensity as shown in panel H. The relative callose staining intensity was set as 1 in the samples of EV (n = 8). Data information: In I), the box boundaries indicate the upper (25th percentile) and lower (75th percentile) quartiles, whiskers indicate minimum and maximum values, central band indicates the median. Letters in the chart denote statistically significant differences among groups according to the Duncan's multiple range test (p < 0.05).