1 1 Supplementary Information 1 Title: Oocytes

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Title: Oocytes express an endogenous red fluorescent protein in a stony coral, Euphyllia. 2 ... millepora (AAU06854), A. hyacinthus (AAU06855), Discosoma sp.

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Supplementary Information

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Title: Oocytes express an endogenous red fluorescent protein in a stony coral, Euphyllia

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ancora: a potential involvement in coral oogenesis.

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Authors: Shinya Shikina, Yi-Ling Chiu, Yi-Jou Chung, Chieh-Jhen Chen, Yan-Horn

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Lee, and Ching-Fong Chang

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Supplementary Table S1, Table S2, Figures, and Figure Legends

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Supplementary Fig. 1

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Deduced amino acid sequences of clone #2-6 (EaRFP) and phylogenetic tree

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comparing with RFP sequences from other anthozoans in Cnidaria. (a) The deduced

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amino acid sequences of clone #2-6 (EaRFP). Black box, chromophore-forming residues.

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Sequence data are available from GenBank, under accession number KT452623. (b) A

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phylogenetic tree comparing the amino acid sequences of RFP from various anthozoans.

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The number at each node represents the bootstrap probability (%); the branches shown

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correspond to values of 50 % and higher. The corresponding GenBank accession

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numbers of the proteins analyzed are as follows: Acropora aculeus (AAU06856), A.

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millepora (AAU06854), A. hyacinthus (AAU06855), Discosoma sp. SSAL-2000

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(AAG16224), Discosoma sp. (AAF03369), Porites porites (ABB17953), Echinopora

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forskaliana (ACD13196), Euphyllia ancora (KT452623), Ricordea florida (AAK71339),

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Montastraea

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Trachyphyllia geoffroyi (BAC20344), Catalaphyllia jardinei (ABN41777), Scolymia

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cubensis (AAU06843), Lobophyllia hemprichii (AAV54099), Mycedium elephantotus

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(ABB17959), Verrillofungia concinn (BAD24721), A. millepora (AAU06852),

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Zoanthus sp. (AAL23574), Montipora efflorescens (ABB17952), Cerianthus sp.

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(AAP55761), Entacmaea quadricolor (AAN05449).

cavernosa

(AAO61598),

Echinophyllia

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echinata

(ABB17960),

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Supplementary Fig. 2

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Fluorescence microscopic observation of the isolated E. ancora tissues. a-d) Isolated

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tentacles (Ten), e-h) Isolated ovary (Ov), i-l) Isolated mesenterial filament (Mf). a, e,

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and i) Bright view. b, f, and j) U-MWIG 2 (RFP) filter view. c, g, and k) U-MWIB 2

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(GFP) filter view. d, h, and l) U-MWU2 (CFP) filter view. All bars = 500 µm.

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Supplementary Fig. 3

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Fluorescence microscopic observation of EaRFP in the ovarian tissues with

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mesenterial filaments. (a,b) The sample collected in August (Aug). (c,d) The sample

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collected in December (Dec). (e,f) The sample collected in March (Mar). a, c, and e:

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bright views; b, d, and f: U-MWIG 2 (RFP) filter-fluorescent views. All bars = 500 µm.

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Supplementary Fig. 4

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Fluorescence microscopic observation of EaRFP in the released egg and the

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embryos. (a,b) The released egg. (c,d) The planula larva (e,f) The larva after settlement

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and metamorphosis. a, c, and e: bright views; b, d, and f: U-MWIG 2 (RFP)

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filter-fluorescent views. All bars = 500 µm.

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