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. 2020 Oct 17;9(10):1383. doi: 10.3390/plants9101383

Figure 2.

Figure 2

Rice RLCK (OsRLCK311) interacts with an aquaporin protein AtPIP2;1 in vivo. (A) AtPIP2;1 interacts with both ΔN and full-length (FL) through co-immunoprecipitation and identified by MS. (B) Co-immunoprecipitation of ΔN or FL with AtPIP2;1 by transient expression in Nicotiana benthamiana, identified by western blots. The proteins extracted from tobacco leaves infiltrated with both ΔN-GFP/FL-GFP (green arrows) and AtPIP2;1-RFP (red arrows) were co-immunoprecipitated (IP) with anti-GFP antibodies and blotted with anti-GFP or anti-red fluorescent protein (RFP) antibodies. Free RFP is marked in white arrows. ‘’Input’’ stands for protein extracts before immunoprepicitaton. (C) Bimolecular fluorescence complementation (BiFC) analysis of the interaction between ΔN or FL with AtPIP2;1 by transient expression in Nicotiana benthamiana. VenusN was fused at the C-terminus of ΔN and FL, while SCFPC was fused to the C-terminus of AtPIP2;1 and OsBIN2. Infiltration with OsBIN2-SCFPC were used as negative controls. AtPIP2;1-mcherry was used as a plasma membrane marker. Bar = 200 μM.