Supplemental Data

Files in this Data Supplement:

• Supplemental Figure 1 - Verification of the T-DNA insertions in shb1-D and shb1 with PCR. (A) SHB1 gene structure. Black boxes represent the 5’- and 3’-untranslated regions. White boxes indicate exons and lines indicate introns. (B). The T-DNA insertion was verified in shb1-Dby PCR analysis with a left border T-DNA primer, lb, and a gene-specific primer, 701. Primer pairs of lb and 701 generated a PCR product only in shb1-Dbut not in Ws wild type. In contrast, two gene-specific primers, X5 and 701, generated a PCR product only in wild type but not in shb1-D.The T-DNA insertion was also verified in the homozygous shb1 lines by PCR analysis using a left border T-DNA primer, lb1, and a gene-specific primer, X3. Fifty pg genomic DNA was used for each PCR reaction.
• Supplemental Figure 2 - Double mutant analysis of shb1-D with phyA-211 and crysunder blue light. Hypocotyl growth responses of Ws/Col lines, shb1-D/phyA-211 lines, shb1-D/cry1 lines, and shb1-D/cry2 lines to 0.6 or 6 μmol/m²/s blue light (Blue-0.6 or Blue-6) for 4 days. Data are presented as means plus or minus standard errors.
• Supplemental Figure 3 - Phylogenetic analysis of SHB1 with its homologues. Phylogenetic tree of SHB1 (gi7487574) with SHB1 homologues from various organisms. This alignment only shows the 3 closest homologues in the PHO1 family: PHO1 H4 (SHB1), PHO1 H5 (gi3548806), and PHO1 H6 (gi 3548805) according to Wang et al. (2004). Bootstrap values are shown at the tree nodes.
• Supplemental Figure 4 - The actual alignment of SHB1 with its homologues. Asterisks indicate positions with a single fully conserved residue. Double dots indicate that one of the following strong groups is fully conserved. Single dots indicate that one of the following weaker groups is fully conserved.
• Supplemental Figure 5 - Sequence alignment of SHB1 with yeast SYG1 protein. Consensus sequence of SPX domain (A) and EXS domain (B) derived from SHB1 and SHB1 homologues, and alignment of SHB1 with SYG1 protein sequence. Highly conserved residues are indicated by shaded reverse contrast, and asterisks indicate similar residues.
• Supplemental Figure 6 - RNA gel blot analysis of CAB3 and CHS expression in shb1/hfr1-201 double mutant. Total RNA was isolated from 5-day old dark-grown Col, shb1, hfr1-201, and shb1/hfr1-201 seedlings that received no light treatment (D) or were treated for 3 hours with blue light (B) under intensities specified in Figure 1A.