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. 2019 Mar 5;17(9):1736–1750. doi: 10.1111/pbi.13096

Figure 2.

Figure 2

Functional in planta complementation of the eif4e1 knock‐out by the five constructed alleles eIF4E1 W69L , eIF4E1 T80 DS 81D , eIF4E1 S84A , eIF4E1 G114R and eIF4E1 N176K . (a) Four‐week‐old plants showing different bolting times. (b) Bolting time in days after sowing assayed on at least 16 plants per genotype. Results for each mutant genotype are pooled from at least two independent lines. Significant differences compared with the wild type, calculated by a Kruskal–Wallis statistical test, are indicated by asterisks for P < 0.05 (*), P < 0.01 (**), P < 0.001 (***) or P < 0.0001 (****). (c) Biochemical assay of cap affinity for the five mutated proteins produced. Total protein extracts were Ponceau stained (bottom panel) and exposed to antibodies directed against actin protein (middle panel). Proteins eluted after passing through a cap‐analogue affinity column were exposed to antibodies directed against the Arabidopsis eIF4E1 protein (top panel). Both experiments were repeated at least twice.