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. 2021 Jan 11;17(1):e1008770. doi: 10.1371/journal.ppat.1008770

Fig 5. βC1 interferes with the dimerization of PIFs.

Fig 5

(A) Modified BiFC competition assays. The EYFP fluorescence was detected after co-expression of GUS + AtPIF3-cEYFP + nEYFP-AtPIF3 (GUS), βC1 + AtPIF3-cEYFP + nEYFP-AtPIF3 (βC1), or GUS + AtPIF3-cEYFP + nEYFP-AtPIF4, βC1 + AtPIF3-cEYFP + nEYFP-AtPIF4. Scale bars = 50 μm. (B-C) Average numbers of EYFP fluorescence show effects of βC1 on the formation of AtPIF3-AtPIF3 homodimers (B) and AtPIF3-AtPIF4 heterodimers (C). Values are mean ± SD (n = 8) (**, P< 0.01; Student’s t-test). (D-E) GST pull-down protein competition assays. The indicated protein amount of His-βC1 or GST was mixed with 2 μg of GST-AtPIF4 and pulled down by 2 μg of MBP-AtPIF4 (D) or 2 μg of MBP-AtPIF3 (E). Immunoblots were performed using anti-GST antibody to detect the associated proteins. Membranes were stained with Coomassie brilliant blue to monitor input protein amount.