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. 2021 Nov 24;49(22):12912–12928. doi: 10.1093/nar/gkab1138

Figure 1.

Figure 1.

Over-expression of wild type ath-pri-MIR168a in transient and transgenic systems. (A) Schematic representation of the AGO1-sensor construct. AGO1-derived part of the sensor is indicated with red, miR168 target site is marked with blue rectangle. (B) Transient over-expression of ath-pri-MIR168a (MIR168a on figures) precursor fragment in the presence of AGO1-sensor. Left panel shows the GFP fluorescence in leaves after co-infiltration of AGO1-sensor construct containing Agrobacteria and either empty vector (pGreen0029) or MIR168a. Right panels indicate the RNA levels of miR168 and the protein levels of the AGO1-sensor in the infiltrated patches. AGO1 part of the sensor fusion protein was detected with antibody raised against ath-AGO1. For the northern blot U6, while for the western blot BiP (Lumenal binding proteins) and Ponceau staining were used as loading controls. (C) Phenotypes of nine weeks old MIR168a precursor fragment over-expressing transgenic lines. Right panels show miR168, miR159 and AGO1 levels in over-expressing and control plants. U6, BiP and Ponceau staining were used as loading controls, respectively. (D) MiR168 distribution among gel-filtration fractions of wild type A. thaliana Columbia (col) and MIR168a over-expressing plants. High molecular weight (HMW) RISCs were detected with AGO1 western and miR159 northern blots. Even fractions were used for protein extraction, odd fractions for RNA purification. Black triangles show positions of known molecular weight markers amongst gel-filtration fractions.