1. Volume 9, Issue 12, Pages 1671-1674 (December 2016)
Dual-Source Nuclear Monomers of UV-B Light Receptor Direct Photomorphogenesis in Arabidopsis 
Chongzhen Qian, Weiwei Mao, Yan Liu, Hui Ren, On Sun Lau, Xinhao Ouyang, Xi Huang 
Molecular Plant 
Volume 9, Issue 12, Pages (December 2016)
DOI: /j.molp
Copyright © 2016 The Author Terms and Conditions

2. Figure 1 Nuclear UVR8 Monomers Direct UV-B Signaling.
(A) Nuclear fractionation analysis of the subcellular distribution and conformational status of YFP-UVR8WT in 4-day-old YFP-UVR8WT/uvr8-6 seedlings grown under −UV-B and +UV-B. PEPC and H3 were used as cytosolic and nuclear markers, respectively. YFP-UVR8d, YFP-UVR8 dimer; YFP-UVR8m, YFP-UVR8 monomer.
(B) Nuclear fractionation analysis of the subcellular distribution and conformational status of YFP-UVR8 variant proteins in 4-day-old YFP-UVR8WT/uvr8-6, YFP-UVR8W285F/uvr8-6 and YFP-UVR8R338A/uvr8-6 seedlings grown under −UV-B and +UV-B. PEPC and H3 were used as cytosolic and nuclear markers, respectively. The asterisk indicates an unspecific cross-reactive band. YFP-UVR8d, YFP-UVR8 dimer; YFP-UVR8m, YFP-UVR8 monomer.
(C) Nuclear translocation analysis of YFP-UVR8 variant proteins. The cytosolic fractions were isolated from 4-day-old YFP-UVR8WT/uvr8-6, YFP-UVR8W285F/uvr8-6 or YFP-UVR8R338A/uvr8-6 seedlings grown under –UV-B. The nuclear fraction was isolated from 4-day-old uvr8-6 seedlings grown under –UV-B. The cytosolic and nuclear fractions were incubated in different combinations under –UV-B or +UV-B for the indicated period. C, cytosolic fraction. N, nuclear fraction. PEPC and H3 were used as cytosolic and nuclear markers, respectively. The asterisk indicates an unspecific cross-reactive band.
(D) Hypocotyl length of 4-day-old wild-type (Col), uvr8-6, YFP-GR-UVR8WT/uvr8-6, YFP-GR-UVR8W285F/uvr8-6, and YFP-GR-UVR8R338A/uvr8-6 seedlings treated or untreated with 20 μM DEX under −UV-B and +UV-B. Error bars represent SD of three biological replicates.
(E) In vivo co-immunoprecipitation assays using 4-day-old YFP-GR-UVR8WT/uvr8-6, YFP-GR-UVR8W285F/uvr8-6, and YFP-GR-UVR8R338A/uvr8-6 seedlings treated or untreated with 20 μM DEX under −UV-B and +UV-B by anti-GFP antibodies. Immunoblot analysis was performed by anti-GFP and anti-COP1 antibodies. Anti-RPN6 was used as an un-immunoprecipitated and loading control. The asterisk indicates an unspecific cross-reactive band.
(F) Nuclear fractionation analysis of the subcellular distribution and conformational status of 4-day-old YFP-UVR8WT/uvr8-6, YFP-UVR8WT/cop1-4, and YFP-UVR8WT/rup1-1 rup2-1 seedlings grown under −UV-B and +UV-B. YFP-UVR8d, YFP-UVR8 dimer; YFP-UVR8m, YFP-UVR8 monomer. PEPC and H3 were used as cytosolic and nuclear markers, respectively. The asterisk indicates an unspecific cross-reactive band.
(G) Nuclear translocation analysis of YFP-UVR8WT in cop1-4 and rup1-1 rup2-1. The isolated cytosolic and nuclear fractions, respectively, were incubated in different combinations under −UV-B or +UV-B for the indicated period. C, cytosolic fraction. N, nuclear fraction. YFP-UVR8d, YFP-UVR8 dimer; YFP-UVR8m, YFP-UVR8 monomer. PEPC and H3 were used as cytosolic and nuclear markers, respectively.
Molecular Plant 2016 9, DOI: ( /j.molp )
Copyright © 2016 The Author Terms and Conditions