HY5 Negatively Regulates FHY1 and FHL Transcript Levels in Vivo.
(A) RNA gel blot analysis showing FHY1 and FHL mRNA levels in 4-d-old wild-type (WT) and hy5 mutant seedlings grown in darkness (D) or continuous FR, R, and B light conditions. Ethidium bromide staining showing rRNA was used as the loading control.
(B) and (C) Real-time qRT-PCR analysis showing FHY1 and FHL transcript levels in wild-type and hy5 mutant seedlings. Error bars represent sd of triplicate experiments. *P < 0.05 and **P < 0.01 (Student’s t test) for the indicated pair of seedlings.
(D) and (E) Changes in FHY1 and FHL transcript levels in wild-type and hy5 mutant seedlings grown in darkness for 4 d and then transferred to FR light for various time periods. The expression levels in dark-grown seedlings were set as 1. Error bars represent sd of triplicate experiments.
(F) and (G) Immunoblots showing the changes of HY5 (F) and FHY3 (G) protein levels in wild-type seedlings grown in darkness for 4 d and then subjected to FR light treatment for various time periods. The mutant plants (hy5-215 and fhy3-1, respectively) were included as negative controls for immunoblots. Asterisk in (F) indicates a band cross-reacting with HY5 antibody. Anti-RPT5 was used as a sample loading control.