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. 2016 May 4;171(2):773–787. doi: 10.1104/pp.16.00335

Figure 4.

Figure 4.

Specific in vivo interaction between BIN2 and the BIN2249-257 APR. A, BiFC assay of BIN2249-257NFT-cGFP coexpressed with different nGFP-tagged ASKs and BIN2249-257NFT-nGFP in tobacco leaves, 3 d after infiltration. In the last panel, the interaction between BIN2249-257NFT-cGFP and MUTE is also shown as a negative control. Bars = 50 μm. B, Coimmunoprecipitation in tobacco leaves of BIN2-HA with different BIN2249-257 SABs after coexpression for 3 d. Booster (B)-GFP, free GFP, BIN2-HA, mock (not infiltrated leaf), and BIN2P249-257NFT-GFP are included as negative controls. Proteins were detected with anti-HA and anti-GFP antibodies. C, Coimmunoprecipitation of BIN2249-257NFT-GFP with MUTE-GS or GS-MUTE proteins coproduced for 3 d as in B. GFP, MUTE-GS, GS-MUTE, and mock are included as negative controls; the GS tag (consisting of a protein G tag and a streptavidin-binding peptide) reacts with the antiperoxidase (PAP) antibody. Anti-PAP and anti-GFP antibodies were used for protein detection.