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. 2021 Jan 16;22(2):870. doi: 10.3390/ijms22020870

Figure 3.

Figure 3

Interaction between COI1a and JAZ15. (A) Y2H assay to screen the interaction between all COIs and JAZs proteins. The top panel shows no interaction between any COI and JAZ without COR supplemented in the medium. Bottom panel shows the interaction in the medium containing 20 µM of COR. Red arrows indicate the interaction between COI1a and JAZs. (B) Split-luciferase assay confirming the interactions between COI1a and selected JAZs. The panel pictures in a, b and c represent different JAZ members to be tested with COI1a, and d is the schematic diagram for different combination of the vectors. (C) Pull-down assay confirming the interaction between COI1a and JAZ15. The assay was conducted using 2 µg COI-MBP and JAZ-GST fusion protein each reaction in the presence of 20 µL GST beads, with 100 nM coronatine. The bands marked in red triangle in top panel shows the signal identified by MBP antibody, and the bands marked in red star in bottom panel shows the input of JAZ-GST fusion protein and GST protein using GST antibody. (D) Interaction between different ZmJAZs and ZmCOI1a point mutation in the presence of 20 µM of COR. (E) ZmJAZ15 point mutants interacting with ZmCOI1a in the presence of 20 µM of COR.