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SupplementaryInformation

Venom gland transcriptome analyses of two freshwater

stingrays(Myliobatiformes:Potamotrygonidae)fromBrazil

NelsonGomesdeOliveiraJúnior,GabrieldaRochaFernandes,MarlonHenrique

Cardoso,FabrícioF.Costa,ElizabetedeSouzaCândido,DomingosGarroneNeto,

MárciaRenataMortari,ElisabethFerroniSchwartz,OctávioLuizFranco,Sérgio

AmorimdeAlencar

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SupplementaryFigures:

FigureS1:TranscriptlengthhistogramsoftheP.amandaeandP.falkneri

Trinity-assembledcontigs.

The length distribution of the P. amandae (blue) and P. falkneri (red) contigs

revealsthatamajorproportionoftheassembledcontigsrangebetween200to

599bpinlength.

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FigureS2:E-valuedistributionofthetopBLASTxhits.

Sequenceswithe-valueequalto0arerepresentedintherightpeak.Thecut-off

usedwas1e-5.P.amandaeandP.falknericontigsarerepresentedinblueandred,

respectively.

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FigureS3:Lysinedegradationpathway.

FigureS4:Arginineandprolinemetabolismpathway.

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FigureS5:Cysteineandmethioninemetabolismpathway.

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FigureS6:Glycine,serineandthreoninemetabolismpathway.

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FigureS7:Citratecycle(TCAcycle)pathway.

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FigureS8:Glycolysis/Gluconeogenesispathway.

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FigureS9:Pyruvatemetabolismpathway.

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FigureS10:Fattyaciddegradationpathway.

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FigureS11:Sphingolipidmetabolismpathway.

FigureS12:Glycerophospholipidmetabolismpathway.

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Figure S13: Clustal Omega alignment of hyaluronidase proteins from a

diversesetoforganisms.

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Figure S14: Summary of the P. amandae and P. falkneri de novowhole

transcriptomeanalysisworkflow.

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Supplementarytables:

Table S1. The 25 most abundant InterPro entries encountered in the P.

amandaeassembledtranscriptome.

InterProAccession InterProDescription CountsIPR027417 P-loopcontainingnucleosidetriphosphatehydrolase 1168IPR011009 Proteinkinase-likedomain 746IPR015880 Zincfinger,C2H2-like 710IPR013083 Zincfinger,RING/FYVE/PHD-type 690IPR000719 Proteinkinasedomain 661IPR007087 Zincfinger,C2H2 642IPR016024 Armadillo-typefold 587IPR013783 Immunoglobulin-likefold 549IPR011993 Pleckstrinhomology-likedomain 540IPR015943 WD40/YVTNrepeat-like-containingdomain 526IPR013087 ZincfingerC2H2-type/integraseDNA-bindingdomain 508IPR017986 WD40-repeat-containingdomain 479IPR001841 Zincfinger,RING-type 477IPR012677 Nucleotide-bindingalpha-betaplaitdomain 422IPR001849 Pleckstrinhomologydomain 419IPR000504 RNArecognitionmotifdomain 390IPR002290 Serine/threonine/dualspecificityproteinkinase,catalyticdomain 370IPR013320 ConcanavalinA-likelectin/glucanasedomain 327IPR011989 Armadillo-likehelical 324IPR007110 Immunoglobulin-likedomain 320IPR001452 SH3domain 306IPR011992 EF-handdomainpair 263IPR029071 Ubiquitin-relateddomain 253IPR000008 C2domain 229IPR011011 Zincfinger,FYVE/PHD-type 228

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Table S2. The 25 most abundant InterPro entries encountered in the P.

falkneriassembledtranscriptome.

InterProAccession InterProDescription CountsIPR027417 P-loopcontainingnucleosidetriphosphatehydrolase 1076IPR013083 Zincfinger,RING/FYVE/PHD-type 679IPR011009 Proteinkinase-likedomain 674IPR000719 Proteinkinasedomain 602IPR016024 Armadillo-typefold 570IPR015880 Zincfinger,C2H2-like 549IPR011993 Pleckstrinhomology-likedomain 519IPR007087 Zincfinger,C2H2 495IPR001841 Zincfinger,RING-type 475IPR015943 WD40/YVTNrepeat-like-containingdomain 454IPR013783 Immunoglobulin-likefold 452IPR017986 WD40-repeat-containingdomain 417IPR001849 Pleckstrinhomologydomain 396IPR013087 ZincfingerC2H2-type/integraseDNA-bindingdomain 379IPR012677 Nucleotide-bindingalpha-betaplaitdomain 364IPR000504 RNArecognitionmotifdomain 329IPR002290 Serine/threonine/dualspecificityproteinkinase,catalyticdomain 325IPR011989 Armadillo-likehelical 319IPR001452 SH3domain 305IPR013320 ConcanavalinA-likelectin/glucanasedomain 281IPR007110 Immunoglobulin-likedomain 265IPR011992 EF-handdomainpair 252IPR029071 Ubiquitin-relateddomain 243IPR011011 Zincfinger,FYVE/PHD-type 235IPR000008 C2domain 216

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Table S3: Theoretical physicochemical parameters obtained for the

modeledproteins.

Models PI Disulfidebonds SASA(Å2) Helix(%) b-sheet(%) Loop(%)

PhospholipaseA2

P.falkneri 9.19 7 8025.41 38.4 8.8 52.8

P.amandae 8.72 7 7766.59 38.4 8.8 52.8

Hyaluronidase

P.falkneri 7.76 2 15860.55 38.6 15.4 46.0

P.amandae 7.76 2 15906.31 38.6 15.4 46.0

CRISP

P.falkneri 7.82 0 9001.23 31.5 17.8 50.7

P.amandae 9.1 0 12119.23 22.4 10.9 66.7

L-aminoacidoxidase

P.falkneri 8.53 0 27563.87 28.6 16.7 54.7

P.amandae 8.43 0 28589.10 31.0 21.0 46.0

Serineprotease

P.falkneri 9.54 4 13587.19 4.7 39.5 55.8

P.amandae 5.54 3 12570.19 1.6 44.4 54.0

VEGFA

P.falkneri 7.31 3 9219.11 7.5 38.3 54.2

Metalloproteinase

P.amandae 5.81 1 9744.49 36 21 41

PI:Theoreticalisoelectricpoint;SASA:solvent-accessiblesurfacearea;Å:Angstrom.

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Table S4:UniprotKBacessionnumbersof theprotein sequences selected

formultiplesequencealignment

UniProtKBaccessionnumber SpeciesXP_013054655 AnsercygnoidesdomesticusXP_008943867 MeropsnubicusXP_005305767 ChrysemyspictabelliiXP_009685663 StruthiocamelusaustralisXP_010213407 TinamusguttatusXP_012678206 ClupeaharengusXP_007896746 CallorhinchusmiliiAAI04791 HomosapiensAAO74499 SynanceiahorridaABI33937 EchisocellatusABI33938 CerastescerastesABI33941 EchispyramidumleakeyiABI33944 BitisarietansACI32917 SalmosalarBAJ54082 SynanceiaverrucosaBAM66299 PteroisantennataBAM66300 PteroisvolitansEMC77798 ColumbaliviaEOB06026 AnasplatyrhynchosNP_001074140 DaniorerioNP_001094250 RattusnorvegicusNP_001119986 XenopustropicalisNP_001295181 GallusgallusXP_001370718 MonodelphisdomesticaXP_001502439 EquuscaballusXP_002190553 TaeniopygiaguttataXP_002752104 CallithrixjacchusXP_002818436 PongoabeliiXP_003217668 AnoliscarolinensisXP_003449275 OreochromisniloticusXP_003967307 TakifugurubripesXP_005810535 XiphophorusmaculatusXP_006012182 LatimeriachalumnaeXP_006024588 AlligatorsinensisXP_006122657 PelodiscussinensisXP_006268718 AlligatormississippiensisXP_006633469 LepisosteusoculatusEMP32607 CheloniamydasXP_007246617 AstyanaxmexicanusXP_527872 Pantroglodytes