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Fig 1.

Biofluorescent Chlopsidae (lower right) photographed in Little Cayman Island.

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Fig 2.

Kaupichthys hyoproroides collected in Bahamas and used in this study.

A) White light; B) Green fluorescence.

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Fig 3.

Details of green fluorescence of Kaupichthys sp.

A) Head fluorescence, B) Cross-section through demonstrating fluorescence in the musculature. C) Close up of skin revealing dark pigmented regions interspersed with fluorescence arising from both the skin and internal musculature.

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Fig 4.

Native gel of tissue homogenate from Kaupichthys hyoproroides.

A) Coomassie stained gel; B) Fluorescent bands imaged under illumination with blue light.

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Table 1.

Summary statistics for individual assemblies of Kaupichthys hyoproroides and Kaupichthys n. sp. (Chlopsid I and Chlopsid II, respectively).

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Table 2.

Summary statistics of read counts and coverage of Kaupichthys hyoproroides.

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Fig 5.

Sequence alignment of fluorescent FABPs from eels with a non-fluorescent FABP from Kaupichthys hyoproroides (Chlopsid NFP) and human brain FABP-7.

Residues highlighted in blue show areas of homology. The GPP sequence motif is highlighted in red.

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Fig 6.

Excitation/emission spectra of Chlopsid FP I and UnaG.

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Fig 7.

Phylogenetic tree generated by Maximum likelihood analysis in RaxML Blackbox.

See text for details of analysis. The arrows in the tree indicate potential branches where duplications occur to explain the paralog patterns in the gene family. The purple circles indicate the two branches where significant dN/dS skew occurs, and the number inside of the circle refers to the magnitude of the dN/dS skew.

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Fig 8.

Diagram showing the alignment of FABP’s and structure of UnaG.

A) Alignment of human brain FABP-7, UnaG and Chlopsid FP I. Red rectangles show residues that are inserted in the eel FP. Green rectangles show residues under dN/dS skew. Grey rectangles indicate polarity index. Blue highlights secondary structure factors. Pink highlights volume. Light green highlights refractivity/heat capacity and purple highlights charge/iso-electric point. When multiple colors appear at a site, this means that more than a single method detected changes or skew at those positions. B) Crystal structure of UnaG (pdb 4I3B) showing areas that are inserted in the eel FP as well as green showing residues under dN/dS skew.

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Table 3.

Table of properties for fluorescent proteins.

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Fig 9.

Two photon image of eel fluorescence.

UnaG distribution in HEK293 cells (top panel), two photon localization of Chlopsid FP I in HEK293 cells (bottom panel) and confocal images of CiVSP-UnaG fusions in HEK293 cells.

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