The transactivation assay of BnaWRKY20 and -53 and their interaction domains with BnaMPKs. Three tests (lanes 1–3) used to determine the interactors of BnaMPKs were cell grown on the nonselective (SD-LT [lane 1] or selective (SD-LTHA [lane 2]) followed by β-galactosidase assay (lane 3). Control tests were BnaMPK-pGBKT7 with empty pGADT7 vectors. (A) Transactivation domain analysis of BnaWRKY20; (B) The interaction assay of BnaWRKY20 as well as it truncated versions with BnaMPK3/6/19. For BnaWRKY20, the peptide with amino acids (AA) 29-77 is the D-domain. D1, AA1-237; D2, AA1-255; D3, AA231-532; (C) Transactivation assay of BnaWRKY53 as well as interaction of its full-length and truncated versions with BnaMPK1/2. For BnaWRKY53, D1, AA1-51; D2, AA1-68; D3, AA44-316; TAD, AA52-60.