Octa probe d 8

Sample preparation for SDS-PAGE was performed by adding Laemmli Sample Buffer. Protein samples were heated at 70 C for 7 min, subjected to 8% polyacrylamide gel electrophoresis and transferred to polyvinylidene fluoride (PVDF) Immobilon-P membrane (Millipore) by the semi-dry transfer system (Bio-Rad). Membranes were blocked with 5% non-fat dry milk in TBST for 60 min and then incubated with rabbit polyclonal primary antibodies diluted as follows, 1:100 OctA-Probe (D-8), 1:2000 anti-GFP (Santa Cruz Biotechnology), 1:1000 anti-GFP (Cell Signaling Technology) used for immunoblots of Figure S2, 1:3000 anti-D1 (Zhang et al., 2000) , 1:3000 anti-CYTF (Alt et al., 1983) , 1:10000 anti-CYTB6/F (Alt et al., 1983) , 1:3000 anti-LHCP (Adamska et al., 2001) and 1:700 anti-CP (Barbato et al., 1992) . Immunoblotting of co-immunoprecipitation experiments performed with mouse monoclonal OctA-Probe (H-5) (Santa Cruz Biotechnology) in 1:500 dilution. After three washes with TBST, the blots were incubated with 1:10000 diluted goat anti-rabbit IgG-HRP or 1:3000 diluted goat anti-mouse IgG-HRP secondary antibody (Santa Cruz Biotechnology). Western blot detection was performed with chemiluminescence luminol reagent (Santa Cruz Biotechnology) by exposure to UltraCruz blue autoradiography film. Western blots were performed in triplicates.