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Supplemental Figure 1. The REVOLUTA M-WD40 subdomain shows similarities to the WD40 domain. Multiple sequence alignment of the Arabidopsis REV M-WD40 subdomain, the Thermomonospora curvata PKWA WD40 domain and the WD40 domain 50% consensus sequence as annotated by the Simple Modular Architecture Research Tool (Schultz et al., 1998). Amino acid grouping: s (A,C,D,G,N,P,S,T,V), p (C,D,E,H,K,N,Q,R,S,T), l (I,L,V), u (A,G,S), a (F,H,W,Y), o (S,T), h (A,C,F,G,H,I,K,L,M,R,T,V,W,Y), + (H,K,R). Schultz, J., Milpetz, F., Bork, P., Ponting, C.P. (1998). SMART, a simple modular architecture research tool: identification of signaling domains. Proc Natl Acad Sci USA 95: 5857-64. Supplemental Figure 2. The leucine zipper domain is necessary for REVOLUTA transcriptional activation activity in vivo. (A) Homo-dimerization yeast two hybrid assays. The same proteins were used as bait (fused to GAL4 DNA binding domain) and prey (fused to GAL4 activation domain). (B) GUS expression in leaf abaxial (lower) epidermal cells of tobacco transiently transformed with pZPR3-uidA and 35S-REV* or pZPR3-uidA and 35S-REV*-ΔLZ constructs. microRNA resistant REV is designated REV*. REV homo-dimerization through the leucine zipper domain is schematically represented by two strings of leucines (L) touching each other. (C) YFP-fusion assays in tobacco leaf abaxial (lower) epidermal cells transiently transformed. Scale bar, 100 μm. Supplemental Figure 3. Multiple sequence alignment of the Arabidopsis thaliana, Oryza sativa, Ginkgo biloba, Pinus taeda, Psilotum nudum, Physcomitrella patens, and Chlamydomonas reinhardtii MEKHLA domains used to generate the Neighbor-Joining tree in Figure 1C. Supplemental Figure 4. Multiple sequence alignment of the Arabidopsis thaliana, Oryza sativa, Ginkgo biloba, Pinus taeda, Psilotum nudum, and Physcomitrella patens HD-ZIP III proteins used to generate the Neighbor-Joining and Maximum-Parsimony trees built to corroborate the tree in Figure 1C. Supplemental Figure 5. Yeast two hybrid assays. Positive interactions were detected by assaying for β-galactosidase activity at eight and 24 hours. Top row are standards for interaction. -, pDEST32/pDEST22. +, pPC97-RB/pPC86-E2F1. ++, pPC97-CYH2– dDP/pPC86-dE2F. +++, pPC97-Fos/pPC86-Jun. 1, pDEST32-REV/pDEST22-REV. 2, pDEST32-REV-ΔMEKHLA/pDEST22-REV-ΔMEKHLA. 3, pDEST32-REV- ΔS2/pDEST22-REV-ΔS2. 4, pDEST32-REV-ΔS1S2-16aa/pDEST22-REV-ΔS1S2-16aa. 5, pDEST32-REV/pDEST22-PHB. 6, pDEST32-REV-ΔMEKHLA/pDEST22-PHB. pDEST32-REV/pDEST22-PHB-ΔMEKHLA. PHB-ΔMEKHLA. 8, 7, pDEST32-REV-ΔMEKHLA/pDEST22- Sequence name REV REV-S2 REV-PAS REV-MEKHLA REV-HD-LZ REV-HD REV-START-HDSAD REV-START REV-HDSAD REV-MEKHLA REV-PAS REV-MEKHLA-S2 REV-M-WD40 REV-S1S2 REV-LEU REV-AASE PHB PHB-MEKHLA ATHB8 ATHB8-MEKHLA ChlamyMEKHLA GL2 ZPR3 * Relative to A in starting ATG Supplemental Table 1. Sequences used in this study. Sequence coordinates* 1-2529 1-2231 1-2187 1-2058 1-453 1-267 383-2058 383-1134 1111-2058 2059-2529 2188-2529 2059-2231 2059-2187 1-2229 1-266375-2529 1-25062511-2529 1-2559 1-2094 1-2502 1-2049 241-708 1-2244 1-204