(A and B) Western blot analyses of PCH1 (A) and PCHL (B) protein (by GFP antibody) in PCH1OE and PCHLOE plants. The plants grown on MS were first illuminated for 4 h to induce germination. Four-day-old dark-grown seedlings were exposed to light for up to 24 h (both left panels) or first incubated in the light for 4 h and then left in darkness up to 8 h (both right panels). RPT5 was detected as loading control. Experiments were repeated three times with the same results, and a representative experiment is shown.
(C) Bar graph shows quantitative data from the western blots shown above. Error bars indicate SEM (n = 3 biological repeats).
(D) Western blots show that PCH1 (left) and PCHL (right) are degraded slower in cop1–4 mutants compared with wild-type background. Four-day-old dark-grown seedlings were illuminated for 4 h (L) or first illuminated for 4 h and then incubated in the dark for another 4 h (D).
(E)YFP-PCH1/PCHL interacts with COP1-HA in vivo. The input and pellet fractions are indicated. Co-IP was carried out using the anti-GFP antibody and then probed with anti-GFP and anti-HA antibodies.