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. 2013 Mar 4;14(3):265–278. doi: 10.1111/mpp.12002

Figure 4.

figure

Self‐interaction of wild‐type and mutants of Chinese wheat mosaic virus (CWMV) cysteine‐rich protein (CRP). (A, C) Bimolecular fluorescence complementation (BiFC) assay: leaves of Nicotiana benthamiana plants were infiltrated with mixtures of Agrobacterium harbouring constructs indicated above and on the left side of the images. Yellow fluorescent protein (YFP) fluorescence was observed using confocal laser scanning microscopy (CLSM) at 3 days after inoculation (dai). Bars, 30 μm. (B, D) Maltose‐binding protein (MBP) pull‐down assay: MBP‐CRP, MBP‐C20A, MBP‐L105A or free MBP (MBP) was mixed with glutathione S‐transferase (GST)‐CRP or unfused GST (GST) and an MBP pull‐down was performed (B). MBP‐ΔC40 was mixed with GST‐ΔC40 or unfused GST (GST) and an MBP pull‐down was performed (D). The purified proteins were analysed by Western blot using GST‐ and MBP‐specific antisera. An asterisk marks the signal from the degraded protein.