Extended Data Fig. 7: ΔCTD accumulation, localization and topology.
(A) ΔCTD localization in chloroplast sub-fractions (Thy: thylakoid, TM: thylakoid membrane after separating lumen, Lumen) in low light (LL) and after 6 h cold and high light treatment (cold HL). Samples were loaded at the same protein amount (3 µg proteins). Proteins were separated by SDS-PAGE and analyzed by immunodetection with antibodies against FLAG, PC, Lhcb4 or ATPb. ATPb is shown as loading control. Representative immunoblot from two independent biological experiments is shown. (B) ΔCTD localization and accumulation compared to wild type (Col-0) in low light (LL) and after 6 h cold and high light treatment (cold HL). Samples were loaded at the same protein amount (3 µg proteins). Proteins were separated by SDS-PAGE and analyzed by immunodetection with anti-SOQ1Trx. Star symbol (*) represents nonspecific band detected by the anti-Trx antibody, based on its absence in soq1-1 lumen samples. Representative immunoblot from three independent biological experiments is shown. (C) The topology of ΔCTD (3 µg chlorophyll) in ΔCTD T2-1, analyzed by immunodetection with anti-FLAG antibody. Protease protection assay on thylakoids treated with thermolysin (Th) in the presence or absence of Triton X-100 (Tr). The stroma-exposed PSI subunit D (PsaD) and the lumenal LCNP are shown as controls. Representative immunoblot from two independent biological experiments is shown. (D) Photosynthetic parameters Fo, Fm, Fv/Fm of Col-0, soq1-1 and three individual T2 ΔCTD lines at time 0 of the NPQ kinetics shown in Fig. 5c. Data represent means ± SD (n = 4 individuals).
