Fig. 5: CsHsp70 contains a CaM-binding domain that interacts with CsCaM and also PdCDIE1.

a Schematic representation of primary structure features of CsHsp70. Three 30aa-peptides of CsHsp70, including peptide A, peptide B, and peptide C, were shown. b Y2H assays were performed to check the putative interactions between three peptides of CsHsp70 and CsCaM, PdCDIE1, N-terminal PdCDIE1 (PdCDIE1^N), or C-terminal PdCDIE1 (PdCDIE1^C). SD-2, SD/–Leu/–Trp; SD-4 + X-α-gal, SD/–Ade/–His/–Leu/–Trp containing X-α-gal. c The putative association of CsCaM with three peptides of CsHsp70 was checked by in vitro peptide binding assays. The association of peptide A and CsCaM resulted in a change in the band of CsCaM. d Co-IP assays showed there was no interaction between CsCaM or PdCDIE1 and CsHsp70^ΔCBD (CsHsp70 without its CaM-binding domain). For Co-IP assays, Agrobacterium-mediated transient gene expression in N. benthamiana leaves was performed. e The putative interaction between CsHsp70 and PdCDIE1^N or PdCDIE1^C was checked by in vitro pull-down assays. f The putative association of PdCDIE1^N or PdCDIE1^C with peptide A of CsHsp70 was checked by in vitro peptide binding assays. The association of peptide A and PdCDIE1^N resulted in a change in the band of PdCDIE1^N. In c, f, Ca²⁺ indicates 1 mM CaCl₂, and 5 mM EGTA was added as a specific Ca²⁺ chelator. All these experiments were repeated three times with similar results.