Fig. 4: Effects of state transitions and exogenous Put on the association state of PGR5.

a The BiFC signal of the PGR5a interaction with PsaH, Lhcb3 and PetC was detected after transient expression in tobacco leaves. N. benthamiana plants with the same growth conditions were selected, and leaves at the same position were injected with the same combination of A. tumefaciens. The seedlings were adapted for 3 days followed by red light illumination and dark adaptation for 30 min, after which the YFP signal was detected. b Quantification of YFP fluorescence intensity. c BN-PAGE with immunoblotting of thylakoid membrane complexes. To arrest the thylakoid membranes in state 1, cucumber discs were incubated with the kinase inhibitor staurosporine at 100 nM for 80 min and then in 20 μM DCMU for 10 min; to isolate state 2-arrested thylakoid membranes, cucumber discs were incubated with the phosphatase inhibitor NaF at 100 mM for 50 min and then in 5 μM carbonyl cyanide p-(trifluoromethoxy)phenylhydrazone (Sigma) for 10 min. Exogenous Put at concentrations of 4, 8, and 10 M was incubated together with the cucumber leaf discs for 60min. The total bands separated by BN-PAGE were used as a loading control. d Relative protein content of PGR5. The different letters indicate significant differences between treatments (P < 0.05) according to Tukey’s test