Figure 4.
F. oxysporum5176 infection reduces the cell wall pectin methylation level in roots, which seems to be detected by RFO1.
(A) Determination of the degree of methylesterification of pectins using FTIR in mock- and F. oxysporum5176-infected roots at 4 dpt. Values correspond to means ± SE of three biological replicates, each one containing at least four technical replicates. Data were analyzed by t-test, ∗p < 0.05.
(B) Quantification of dot immunobinding assays from membranes of immobilized plant cell wall fractions from Mock- and F. oxysporum5176-infected WT roots at 4 dpt and probed with JIM5 (anti-dmPectin) or JIM7 (anti-mPectin) antibodies as shown in Supplemental Figure 7A.
(C) Quantification of dot immunobinding assay using root cell wall fractions I and II as described in (B) and F. oxysporum5176 hyphae cell wall fractions probed with RFO1ECD-SNAP recombinant protein as shown in Supplemental Figure 7B.
(B and C) Boxplots: centerlines show the medians; means are marked by +; box limits indicate the 25th and 75th percentiles; whiskers extend to the minimum and maximum. N ≥ 12 from ≥3 independent replicates. For each replicate, data are reported as the percentage that each fraction represents from the total gray value of the blot. Two-way ANOVA with Tukey’s multiple-comparison test, ∗p < 0.05, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.