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. 2020 Oct 19;30(20):4103–4111.e6. doi: 10.1016/j.cub.2020.07.095

Figure 3.

Figure 3

Relations between UCC1 Positioning and Other Components of the Casparian Strips Machinery

(A) Analysis of the spatial distribution of CASP1 and UCC1 at the endodermal cell junctions. Images were generated from the same plant co-expressing CASP1-GFP and mCherry-UCC1 using confocal microscopy. The numbers at the bottom of the figure indicate the number of cells after the onset of elongation. White arrows indicate the central accumulation for CASP1-GFP or mCherry-UCC1. Scale bar, 6 μm.

(B) Histograms showing the frequency distribution (Frequency [%]) of the onset of expression (upper plot, n = 18) and the onset of localization at the Casparian strip of CASP1-GFP and mCherry-UCC1 (lower plot, n = 28).

(C) Maximum intensity projection (left) and surface view (right) of UCC1 immunolocalization (red) at 10 cells after the onset of elongation in WT plants and a collection of Casparian strips mutants: casp1 casp3, esb1, sgn3, esb1 sgn3. White arrows show gaps in the UCC1 localization. Scale bar, 20 μm for the maximum projections and 2 μm the surface views. see also Figure S3A.

(D) Maximum intensity projection of CASP1-GFP localization in cleared root of WT plants and the mutants ucc1.1 and ucc1.1 ucc2.1. Scale bar, 20 μm.

(E) Surface view of the localization of CASP1-GFP in cleared root of WT plants and the mutants: ucc1.1, ucc1.2, and ucc1.1 ucc2.1. Scale bar, 2 μm.

(F) Quantification of normalized pixels intensity (relative pixel intensity; 0–1) across the Casparian strip in plants expressing CASP1-GFP. The plots are showing the intensity profile for individual replicates (n ≥ 10), the mean value (black line), and the 95% confidence interval (gray interval). Each replicate corresponds to the quantification of one picture containing a Casparian strip segment of approximately 25 μm long. The pictures were generated at the 15th cells after the onset of elongation from at least 8 individual plants per genotype. Intensity profiles across the Casparian strip were always measured in the same orientation—from the cortical side toward the pericycle side of the endodermis. Letters indicate statistically significant differences between genotypes for the intensity values comprised between the dashed lines using an ANOVA and Tukey’s test as post hoc analysis (p < 0.01) (see also Figures S3B and S3C).