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. 2020 Dec 3;21(23):9237. doi: 10.3390/ijms21239237

Figure 2.

Figure 2

BiFC assays between arabidopsis GRXS15 and ISCA proteins in arabidopsis leaf protoplasts. Arabidopsis protoplasts obtained from four week-old plantlets were transfected with combinations of two vectors expressing GRXS15 fused to the N-terminal region of YFP (GRXS15-N in panels) and ISCAs cloned upstream of the C-terminal region of YFP (ISCA-C in panels). The YFP fluorescence was recorded 24 h post-transfection by confocal microscopy. All confocal images shown here were captured using a maximum Z-stack intensity projection. Images showing confocal plans without Z-stack intensity projection are shown in Figure S3. Negative controls verifying that none of the proteins expressed alone can restore YFP fluorescence are shown. BiFC results obtained using opposite protein fusion conformations (GRXS15-C coexpressed with ISCA-N) showed less clear-cut results as a strong fluorescence in the cytosol suggested the formation of aggregates was obtained for some combinations. Bars = 10 μm.