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. 2021 Aug 6;12:713544. doi: 10.3389/fpls.2021.713544

Figure 6.

Figure 6

Osj10gBTF3 enters the chloroplast stroma and recruits OsCpn60 to OsPPR676. (A) and (B) Co-IP analysis of OsHSP82 (A) or Osj10gBTF3 (B) and Toc64 or Cpn60 by corresponding antibodies. Co-precipitated proteins were identified by Western blotting. The IgG2a and IgG2b were used as isotype control for RFP and Osj10gBTF3 antibodies, respectively. Ab, antibody; I, input; and IP, immunoprecipitation. Three independent replicates were performed for each experiment. (C) Analysis of the interaction between Osj10gBTF3 and the presumed channel proteins at the inner membrane of chloroplast. The pGADT7-T cells co-transformed with pGBKT7-53 or the pGBKT7-Lam were used as positive or negative controls, respectively. Three independent replicates were performed for the experiment. (D) Co-IP verification analysis of the interaction between Osj10gBTF3 and chloroplast inner membrane protein Tic110. The proteins were extracted from leaves of rice wild-type seedlings. The input is 1/4 of total loading lysate. IgG2b were used as isotype control for Osj10gBTF3 antibodies. I, input; IP, immunoprecipitation. Three independent replicates were performed for the experiment. (E–G) MST was performed to determine the equilibrium dissociation constants (Kd) between Osj10gBTF3, OsCpn60, and OsPPR676. Values represent the mean ± SD, n = 3 independent replicates.