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. 2017 Apr 3;7:45590. doi: 10.1038/srep45590

Figure 7. Interaction between the N-terminus of CWMV replicase and TaHSP70 or NbHSP70 in vivo and vitro.

Figure 7

(A) Visualization of the interaction between Rep1–333 and TaHSP70 or NbHSP70 in N. benthamiana epidermal cells by BiFC assay. N. benthamiana leaves were co-infiltrated with recombinant BiFC vectors containing the constructs indicated above the images. The results were observed 48 h after infiltration. Scale bar, 50 μm. The fluorescent and merged images are depicted in the upper and lower panels, respectively. (B,C) Interaction of CWMV Rep1–333 with TaHSP70 and NbHSP70 proteins in ELISA-based binding assays. Wells of a microtiter plate were coated with 25 pmol of E. coli purified GST-tagged Rep1–333 protein and incubated with increasing amounts of E. coli purified 6×-histidine-tagged TaHSP70 protein (▲) or 6×-histidine-tagged NbHSP70 protein (▼), respectively. Retention of the complex was detected with polyclonal anti-His antibodies. The recombinant 6×-histidine tag was used alone as a control (■). Experiments were repeated three times. Bars represent the standard errors of the means.