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. 2022 Mar 9;11(6):938. doi: 10.3390/cells11060938

Figure 3.

Figure 3

Characterization of β’1β’3-cop double mutants. (A) All the β’1β’3-cop homozygous double mutants show a dwarf phenotype at 7-day-old seedling stage. At later stages, the β’1β’3-cop-1 mutant also showed a dwarf phenotype with smaller rosette leaves, shorter stems and roots and reduced fertility (Supplementary Figure S3). No homozygous lines could be obtained of β’1β’3-cop-2 and β’1β’3-cop-3 mutants as they were only viable as seedlings and failed to develop beyond the seedling stage. White arrows point β’1β’3-cop-2 and β’1β’3-cop-3 homozygous seedlings obtained from seeds of β’1-cop-1/β’1-COP β’3-cop-2/β’3-cop-2 plants and β’1-cop-1/β’1-cop-1 β’3-COP/β’3-cop-3 plants, respectively. (B) RT–qPCR analysis show the expression levels of the three β’-COP genes in β’1β’3-cop double mutants relative to the wild type (Col-0). (C) β’1β’3-cop-1, β’1β’3-cop-2 and β’1β’3-cop-3 mutants show upregulation of the COPII subunit SEC31A gene. Expression of SEC31A and SEC31B was analyzed by RT–qPCR. Total RNA was extracted from 7-day-old seedlings of wild type (Col-0) and homozygous mutants (β’1β’3-cop-2 and β’1β’3-cop-3 were selected by size). The mRNA was analyzed by RT–qPCR with specific primers and normalized to UBQ10 expression (Supplementary Tables S1–S3). Results are from three biological samples and three technical replicates. mRNA levels are expressed as relative expression levels and represent fold changes of mutant over wild type. Values represent mean ± s.e.m. of the three biological samples.