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. 2016 Nov 28;28(12):2916–2936. doi: 10.1105/tpc.16.00609

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Figure 6. — NKD1 and NKD2 Protein BiFC and Co-Pull-Down.

(A) Transient BiFC assays in onion epidermal cells for full-length NKD1 and NKD2 proteins. Vectors used for each BiFC assay are listed to the left of each row. Arrows designate nuclei viewed under differential interference contrast (DIC). mCherry florescence marks transient cells transformation and YFP florescence indicates a positive protein-protein interaction. Control bombardments containing NYFP + NKD1-CYFP or NKD2-NYFP + CYFP did not produce YFP fluorescence. Bars = 100 μm. Additional controls are shown in Supplemental Figure 12.

(B) and (C) Co-pull-down immunoblots for full-length NKD1 and NKD2 proteins (B) and NKD1-ID and NKD2-ID proteins (C). Affinity column and antibody (Ab) used for each assay are indicated at the top of each panel. Total soluble bacterial lysates (inp) were immunoblotted as a positive control for protein expression. The first set of control lanes show no cross-reaction between NKD-GST protein with anti-6xHis antibody or NKD-6xHis protein with anti-GST antibody.