FIGURE 1.

GmDNJ1 is a typical type‐I HSP40. (a) A cartoon depicting the protein structure of GmDNJ1. The various domains were determined by Pfam (El‐Gebali et al., 2019). The nuclear localization signal (NLS) and nuclear export signal (NES) were determined by LOCALIZER (Sperschneider et al., 2017) and NetNES 1.1 Server (la Cour et al., 2004), respectively. (b) Phylogenetic tree of characterized HSP40s. The tree was built with MEGA X (Kumar et al., 2018) using the Maximum Likelihood method and the JTT matrix‐based model with 1,000 bootstrap replications for phylogeny test. Bootstrap values were labeled as percentages on the branches. The accession numbers of the HSP40 protein sequences included in this tree are GmDNJ1: NP_001238341.1; AtJ3: NP_189997.1; AtDjB1: NP_195759.1; AtDjC1: NP_187752.2; HsDNAJA1: NP_001530.1; HsDNAJB1: NP_006136.1; HsDNAJC9: NP_056005.1; ScYdj1: NP_014335.1; ScSis1: NP_014391.1; EcDnaJ: NP_414556.1; OsDjA5: XP_015632121.1; OsDjB7: XP_015637533.1; OsDjC12: EEE55258.1; AtHSP70: NP_001328002.1; and EcDnaK: WP_102804582.1. (c) Subcellular localization study of GmDNJ1. A total of >50 cells were observed for each line with similar results. (d) Luciferase refolding assay for testing GmDNJ1 co‐chaperone activity. The luciferase activity in each reaction was normalized to that containing E. coli DnaJ–DnaK‐GrpE in combination (lane 3), which was set at 100%. Each bar represents the average of at least three technical replicates with the error bar representing standard error