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. 2020 Nov 23;33(1):104–128. doi: 10.1093/plcell/koaa008

Figure 5.

Figure 5

Mutation in ZmABI19 Affects the expression of O2, Pbf1, and zein genes. (A), (C), and (F) RT-qPCR analysis of O2, Pbf1, and zein expression in 16-DAP wild-type and zmabi19-1 endosperms. All expression levels were normalized to that of Actin. Error bars represent SD of three biological replicates. **P < 0.01. (B) Immunoblot analysis of ZmABI19 and O2 proteins in wild-type and zmabi19-1 seeds. The number below each lane indicates the protein levels relative to the values in wild type set as 1, which were all normalized against ACTIN. (D) EMSA of the specific binding of ZmABI19-His to the RY motif in the Pbf1 promoter. The normal and mutated RY motifs are indicated in magenta and green, respectively. Mp, mutated probes; M-Comp, mutated competitor. (E) DLR assay of the transactivation of the Pbf1 promoter by ZmABI19. Shown in the relative ratio of LUC/REN tested in Arabidopsis protoplasts via co-transfecting the reporter plasmid with effector constructs. **P < 0.01. (G) SDS-PAGE analysis of (left) zein and (right) non-zein proteins in wild-type and zmabi19-1 seeds. The size of each zein protein band is indicated beside it. γ50, 50-kD γ-zein; γ27, 27-kD γ-zein; α22, 22-kD α-zein; α19, 19-kD α-zein; γ16, 16-kD γ-zein; β15, 15-kD β-zein; and δ10, 10-kD δ-zein.