Fig. 1
From: PCH1 and PCHL promote photomorphogenesis in plants by controlling phytochrome B dark reversion
Light-activated phyB interacts with PCH1 and PCHL. a, b Y2H detection of interaction of phyB with PCH1 and PCHL. The phyB-GAL4-activation domain (phyB-AD) fusion was co-expressed with GAL4-DNA-binding domain (BD-) fusions of PCH1 or PCHL. a X-Gal filter-lift assay. Yeast cells were lifted from chromophore-supplemented plates pre-incubated for 48 h in either constant darkness (D), red (R), or far-red light (FR). b ONPG assay. Yeast cultures supplemented with chromophore were exposed to red light, to convert phyB to Pfr, or to far-red light for phyB Pr. n = 3; error bars indicate ± s.e.m. c, d PCH1 and PCHL co-purify with phyB from native plant extracts. Four-day-old dark-grown seedlings over-expressing HA-YFP-PCH1 (PCH1ox) c or HA-YFP-PCHL (PCHLox) d in the Col-0 wild type background were either treated with red light (R, 7 μmol m−2 s−1) for 10 min or kept in darkness (D). Native total protein extracts were prepared and used for co-immunoprecipitation (Co-IP) assays. IP was performed using α-GFP antibody. α-phyB and α-GFP antibodies were used to detect endogenous phyB and YFP-tagged PCH1 and PCHL, respectively
