M.Musculusmstn MMQKLQ MYVYIYLFMLIAAGP VDLNEGSE 29

Fig.S1

B.belcheriGDF8/11 -MHMHANDVD-IEVLQWTTSEEVNVNSTRERLSGS------SVDTAVPTTAYG 45

B.floridaeGDF8/11 -MHMHANDVDTVEALQWTTSEDVNVNSTRERPSGS------SVD-AAPTTAYG 45

M.musculusMSTN MMQKLQ------MYVYIYLFMLIAAGP------VDLNEGSE--- 29

R.norvegicusMSTN MIQKPQ------MYVYIYLFVLIAAGP------VDLNEDSE--- 29

H.sapiensMSTN -MQKLQ------LCVYIYLFMLIVAGP------VDLNENSE--- 28

G.gallusMSTN -MQKLA------VYVYIYLFMQIAVDP------VALDGSSQ--- 28

M.gallopavoMSTN ------MQILVHP------VALDGSSQ--- 15

R.norvegicusGDF11 -MVLAA------PLLLGFLLLALELRPRGEAAEGP--AAAAAAAAAAAGVGGERSSR--- 48

M.musculusGDF11 -MVLAA------PLLLGFLLLALELRPRGEAAEGP--AAAAAAAAAAAGVGGERSSR--- 48

H.sapiensGDF11 -MVLAA------PLLLGFLLLALELRPRGEAAEGPAAAAAAAAAAAAAGVGGERSSR--- 50

D.rerioGDF11 -MKRYN------FLLCLTVLISLGLS----GSDEP-----NLFLAPLSEMSSDIGVS--- 41

T.rubripesMSTN1 -MQLSP------SMLHFSLMISLSLVVLSG------QETHQQ--- 29

S.aurataMSTN -MHPSQ------IVLYLSLLIVLGPVVLSE------QETQQQQQQ 32

D.rerioMSTN1 -MHFTQ------VLISLSVLIACGPVGYGD------ITAHQQ--- 29

T.rubripesMSTN2 ------MLVLAVLTVVS-AGFSM------EMNQTS--- 22

S.aurataMSTNb ------MLVFLGLTVLLSAGSSV------EMNQTS--- 23

D.rerioMSTN2 -MFLLF------YLSFWGVLGSQNQNLSTT------TTTTTQ--- 29

: : : :

B.belcheriGDF8/11 -----EEATGPPGETSKCGAARTRGQEREMRVETLKRHILDKLGLKRSPTIPR---NRTL 97

B.floridaeGDF8/11 -----AEATPPPGETSKCGASCTRGQEREMRVETLKRHILDKLGLKRSPTIPR---NRTL 97

M.musculusMSTN ------REENVEKE-GLCNACAWRQNTRYSRIEAIKIQILSKLRLETAPNISKDAIRQLL 82

R.norvegicusMSTN ------REANVEKE-GLCNACAWRQNTRYSRIEAIKIQILSKLRLETAPNISKDAIRQLL 82

H.sapiensMSTN ------QKENVEKE-GLCNACTWRQNTKSSRIEAIKIQILSKLRLETAPNISKDVIRQLL 81

G.gallusMSTN ------PTENAEKD-GLCNACTWRQNTKSSRIEAIKIQILSKLRLEQAPNISRDVIKQLL 81

M.gallopavoMSTN ------PTENAEKD-GLCNACTWRQNTKSSRIEAIKIQILSKLRLEQAPNISRDVIKQLL 68

R.norvegicusGDF11 ------PAPSAAPEPDGCPVCVWRQHSRELRLESIKSQILSKLRLKEAPNISREVVKQLL 102

M.musculusGDF11 ------PAPSAPPEPDGCPVCVWRQHSRELRLESIKSQILSKLRLKEAPNISREVVKQLL 102

H.sapiensGDF11 ------PAPSVAPEPDGCPVCVWRQHSRELRLESIKSQILSKLRLKEAPNISREVVKQLL 104

D.rerioGDF11 ------LFDVDDVESSECSACVWREQSKVLRLETIKSQILSKLRLKQAPNISREVVNQLL 95

T.rubripesMSTN1 ------PPVGSPEDTEQCVTCDVRQHIKTMRLNAIKPQILSKLRMKEAPNISRDTVKQLL 83

S.aurataMSTN QQQQQQPSATSPEDTELCATCEVRQQIKTMRLNAIKSQILSKLRMKEAPNISRDIVKQLL 92

D.rerioMSTN1 ------PSTATEESELCSTCEFRQHSKLMRLHAIKSQILSKLRLKQAPNISRDVVKQLL 82

T.rubripesMSTN2 ------RLLAESGEQCSACDFREHSKQMRLHSIKSQILSILRLEQAPNISRDMIRQLL 74

S.aurataMSTNb ------KLLAESGEQCSACDFREHSKQMRLHSIKSQILSILRLEQAPNISRDMIRQLL 75

D.rerioMSTN2 ------AFVTPGDDNGQCTTCQFRQQSKLLRLHSIKSQILSILRLEQAPNISRDTVKLLL 83

. * .. * : : *:.::* :**. * :: :*.*.: . *

B.belcheriGDF8/11 PRAPPMQSLLDQYGFYPDRSGNIVSAEDITQAFQGDSPASYNYHLVDTERRHYEDSAETE 157

B.floridae GDE8/11 PRAPPMQSLLDQYGFYPDRSGNIVSAEDITQAFQGDSPASYNYHLVDTERRHYEDVAETE 157

M.musculusMSTN PRAPPLRELIDQY------DVQRDDSS-DGSLEDDDYH------ATTE 117

R.norvegicusMSTN PRAPPLRELIDQY------DVQRDDSS-DGSLEDDDYH------ATTE 117

H.sapiensMSTN PKAPPLRELIDQY------DVQRDDSS-DGSLEDDDYH------ATTE 116

G.gallusMSTN PKAPPLQELIDQY------DVQRDDSS-DGSLEDDDYH------ATTE 116

M.gallopavoMSTN PKAPPLQELIDQY------DVQRDDSS-DGSLEDDDYH------ATTE 103

R.norvegicusGDF11 PKAPPLQQILDLH------DFQGDALQPEDFLEEDEYH------ATTE 138

M.musculusGDF11 PKAPPLQQILDLH------DFQGDALQPEDFLEEDEYH------ATTE 138

H.sapiensGDF11 PKAPPLQQILDLH------DFQGDALQPEDFLEEDEYH------ATTE 140

D.rerioGDF11 PKAPPLQQLLDHH------DFQGDASSLEDFIDADEYH------ATTE 131

T.rubripesMSTN1 PKAPPLQQLLDQY------DVLGDDNR-DVVTEEDDEH------AITE 118

S.aurataMSTN PKAPPLQQLLDQY------DVLGDDNR-DVVMEEDDEH------AITE 127

D.rerioMSTN1 PKAPPLQQLLDQY------DVLGDDSK-DGAVEEDDEH------ATTE 117

T.rubripesMSTN2 PKAPPLTQLLDQY------DPR------VEDEDH------ATTE 100

S.aurataMSTNb PKAPPLTQLLDQY------DPR------VEEEDH------ATTE 101

D.rerioMSTN2 PKAPPLQELLDQY------DQNGG------ISEDEEQ------ASSE 112

*:***: .::* : : :: : : : * :*

B.belcheri GDF8/11 TVITVATPP-PVAPFNESG-CCYFKFGRHVSRTKVNKAFLWLYVRAAEDMQSTSIPLKIY 215

B.floridae GDE8/11 TVITVATPP-PVAPFNESG-CCFFKFGRHVSRTKVNKAFLWLHVRAAENMQSTSIPLKIY 215

M.musculusMSTN TIITMPTESDFLMQADGKPKCCFFKFSSKIQYNKVVKAQLWIYLRPVK--TPTTVFVQIL 175

R.norvegicusMSTN TIITMPTESDFLMQADGKPKCCFFKFSSKIQYNKVVKAQLWIYLRAVK--TPTTVFVQIL 175

H.sapiensMSTN TIITMPTESDFLMQVDGKPKCCFFKFSSKIQYNKVVKAQLWIYLRPVE--TPTTVFVQIL 174

G.gallusMSTN TIITMPTESDFLVQMEGKPKCCFFKFSSKIQYNKVVKAQLWIYLRQVQ--KPTTVFVQIL 174

M.gallopavoMSTN TIITMPTESDFLVQMEGKPKCCFFKFSSKIQYNKVVKAQLWIYLRQVQ--KPTTVFVQIL 161

R.norvegicusGDF11 TVISMAQETDPAVQTDGSPLCCHFHFSPKVMFTKVLKAQLWVYLRPVP--RPATVYLQIL 196

M.musculusGDF11 TVISMAQETDPAVQTDGSPLCCHFHFSPKVMFTKVLKAQLWVYLRPVP--RPATVYLQIL 196

H.sapiensGDF11 TVISMAQETDPAVQTDGSPLCCHFHFSPKVMFTKVLKAQLWVYLRPVP--RPATVYLQIL 198

D.rerioGDF11 SVITMASEPEPLVQVDGKPTCCFFKFSPKLMFTKVLKAQLWVYLQPLK--QTSTVYLQIL 189

T.rubripesMSTN1 TIMMMATEPASVVQVNGEPKCCHFSFTQKFQVSRLVRAQLWVHLRPAA--EATTVFLQIS 176

S.aurataMSTN TIMMMATEPEPVVQVDGEPRCCFFSFTQKIQANRIVRAQLWVHLRASD--EANTVFLQIS 185

D.rerioMSTN1 TIMTMATEPDPIVQVDRKPKCCFFSFSPKIQANRIVRAQLWVHLRPAE--EATTVFLQIS 175

T.rubripesMSTN2 TIITMATKPNPIAQ-DALSSCCLLSLSPKIQPKNILRALLWVHLRPAD--TVATVFLQIS 157

S.aurataMSTNb TIITMATKHNPIAQ-DELTSCCLFSLSPKIQPKNILRAQLWVHLRPAD--IVTSVFLQIS 158

D.rerioMSTN2 TIITMATEPQAITQLVGMPKCCMFALSPKILPDSILKALLWIYLRPAE--EPTTVYIQIS 170

::: :. ** : : :. : :* **:::: : :: ::*

B.belcheriGDF8/11 RIAQASSFYT------EPYKVLISSHKITDVKPNEGLWVSYPIRDQVRRWFHNPAENLGI 269

B.floridaeGDF8/11 RIAPASSAYP------EPYKILISSHKITDVKPNEGVWVSYPIRDQVRRWFHNPAENLGI 269

M.musculusMSTN RLIKPMKD------GTRYTGIRSLKLDMS-PGTGIWQSIDVKTVLQNWLKQPESNLGI 226

R.norvegicusMSTN RLIKPMKD------GTRYTGIRSLKLDMS-PGTGIWQSIDVKTVLQNWLKQPESNLGI 226

H.sapiensMSTN RLIKPMKD------GTRYTGIRSLKLDMN-PGTGIWQSIDVKTVLQNWLKQPESNLGI 225

G.gallusMSTN RLIKPMKD------GTRYTGIRSLKLDMN-PGTGIWQSIDVKTVLQNWLKQPESNLGI 225

M.gallopavoMSTN RLIKPMKD------GTRYTGIRSLKLDMN-PGTGIWQSIDVKTVLQNWLKQPESNLGI 212

R.norvegicusGDF11 RLKPLTGEGTAGGGGGGRRHIRIRSLKIELH-SRSGHWQSIDFKQVLHSWFRQPQSNWGI 255

M.musculusGDF11 RLKPLTGEGTAGGGGGGRRHIRIRSLKIELH-SRSGHWQSIDFKQVLHSWFRQPQSNWGI 255

H.sapiensGDF11 RLKPLTGEGTAGGGGGGRRHIRIRSLKIELH-SRSGHWQSIDFKQVLHSWFRQPQSNWGI 257

D.rerioGDF11 RLKPITEQ------GSRHIRIRSLKIELD-SQAGHWQSIDFKHVLQNWFKQPHTNWGI 240

T.rubripesMSTN1 RLMPVTDGN------RHIRIRSLKLDVK-AGVSSWQSIDVKQVLSVWLRQPETNWGI 226

S.aurataMSTN RLMPVTDGN------GHIHIRSLKIDVN-AGVGSWQSIDVKQVLSVWLRQPETNWGI 235

D.rerioMSTN1 RLMPVKDGG------RHR-IRSLKIDVN-AGVTSWQSIDVKQVLTVWLKQPETNRGI 224

T.rubripesMSTN2 RLKPGIEGNN------TRVRVRSLRIDTDTAGAGSWQSVDIKSLLQAWLRQPETNYGI 209

S.aurataMSTNb RLKPGKEGNN------TRIRVRSLKIDTD-AGAGSWQSIDIKSLLQAWLRQPETNYGI 209

D.rerioMSTN2 HLESSSEGN------NHSRIRAQKIDVN-ARTNSWQHIDMKQLLKLWLKQPQSNFGI 220

:: : : :: . * .: : *:::* * **

B.belcheri GDF8/11 VIETS-EAGKDLIVLDPKPGEEPLQPFLVMHTHDPGTVRRKRMAGLNCEQDSNEERCCRY 328

B.floridae GDF8/11 VIETS-EAGKDLIVLDPKPGEEPLQPFLVMHTHDPGTVRRKRMAGLNCEQDSNEERCCRY 328

M.musculusMSTN EIKALDENGHDLAVTFPGPGEDGLNPFLEVKVTD-TPKRSRRDFGLDCDEHSTESRCCRY 285

R.norvegicusMSTN EIKALDENGHDLAVTFPGPGEDGLNPFLEVKVTD-TPKRSRRDFGLDCDEHSTESRCCRY 285

H.sapiensMSTN EIKALDENGHDLAVTFPGPGEDGLNPFLEVKVTD-TPKRSRRDFGLDCDEHSTESRCCRY 284

G.gallusMSTN EIKAFDETGRDLAVTFPGPGEDGLNPFLEVRVTD-TPKRSRRDFGLDCDEHSTESRCCRY 284

M.gallopavoMSTN EIKAFDENGRDLAVTFPGPGEDGLNPFLEVRVTD-TPKRSRRDFGLDCDEHSTESRCCRY 271

R.norvegicusGDF11 EINAFDPSGTDLAVTSLGPGAEGLHPFMELRVLE-NTKRSRRNLGLDCDEHSSESRCCRY 314

M.musculusGDF11 EINAFDPSGTDLAVTSLGPGAEGLHPFMELRVLE-NTKRSRRNLGLDCDEHSSESRCCRY 314

H.sapiensGDF11 EINAFDPSGTDLAVTSLGPGAEGLHPFMELRVLE-NTKRSRRNLGLDCDEHSSESRCCRY 316

D.rerioGDF11 DINAYDESGNDLAVTSLGPGEEGLQPFLEVKILE-TTKRSRRNLGLDCDEHSTESRCCRY 299

T.rubripesMSTN1 EINAFDSRGKDLAVTSTQPGEEGLQPFMEVKISE-GPRRVRRDLGLDCDENSPESRCCRY 285

S.aurataMSTN QINAFDSRGNDLAVTSAEPGEDGLQPFMEVKISE-GPKRVRRDSGLDCDENSPESRCCRY 294

D.rerioMSTN1 EINAYDAKGNDLAVTSTETGEDGLLPFMEVKISE-GPKRIRRDSGLDCDENSSESRCCRY 283

T.rubripesMSTN2 EINAFDSKGEDRAVTSLEPGEEGLQPFIEVKILN-SPKRSRRESGLNCDEESAETRCCRY 268

S.aurataMSTNb EINAYDSKGEDLAVTSAEPGEEGLQPFIEVKILD-NPKRSRRDSGLNCDEESAETRCCRY 268

D.rerioMSTN2 EIKAFDANGNDLAVTSTESGEEGLQPFLEVKISD-TGKRSRRDTGFDCDEHSTESRCCRY 279

*:: * * * .* : * **: :: : * :* *::*::.* * *****

B.belcheri GDF8/11 PLQVDFREFGWDWIIAPNTYQAYYCAGDCPPIFLQKYPHTHLVQQVNKAGSVGPCCTPTK 388

B.floridae GDF8/11 PLQVDFREFGWDWIIAPNTYQAYYCAGDCPPIFLQKYPHTHLVQQVNKAGSVGPCCTPTK 388

M.musculusMSTN PLTVDFEAFGWDWIIAPKRYKANYCSGECEFVFLQKYPHTHLVHQANPRGSAGPCCTPTK 345

R.norvegicusMSTN PLTVDFEAFGWDWIIAPKRYKANYCSGECEFVFLQKYPHTHLVHQANPRGSAGPCCTPTK 345

H.sapiensMSTN PLTVDFEAFGWDWIIAPKRYKANYCSGECEFVFLQKYPHTHLVHQANPRGSAGPCCTPTK 344

G.gallusMSTN PLTVDFEAFGWDWIIAPKRYKANYCSGECEFVFLQKYPHTHLVHQANPRGSAGPCCTPTK 344

M.gallopavoMSTN PLTVDFEAFGWDWIIAPKRYKANYCSGECEFVFLQKYPHTHLVHQANPRGSAGPCCTPTK 331

R.norvegicusGDF11 PLTVDFEAFGWDWIIAPKRYKANYCSGQCEYMFMQKYPHTHLVQQANPRGSAGPCCTPTK 374

M.musculusGDF11 PLTVDFEAFGWDWIIAPKRYKANYCSGQCEYMFMQKYPHTHLVQQANPRGSAGPCCTPTK 374

H.sapiensGDF11 PLTVDFEAFGWDWIIAPKRYKANYCSGQCEYMFMQKYPHTHLVQQANPRGSAGPCCTPTK 376

D.rerioGDF11 PLTVDFEAFGWDWIIAPKRYKANYCSGQCEYMFMQKYPHTHLVQHANPRGSAGPCCTPTK 359

T.rubripesMSTN1 PLTVDFEDFGWDWIIAPKRYKANYCSGECEYMHLQKYPHTHLVNKANPRGTAGPCCTPTK 345

S.aurataMSTN PLTVDFEDFGWDWIIAPKRYKANYCSGECEYMHLQKYPHTHLVNKANPRGSAGPCCTPTK 354

D.rerioMSTN1 PLTVDFEDFGWDWIIAPKRYKANYCSGECDYMYLQKYPHTHLVNKASPRGTAGPCCTPTK 343

T.rubripesMSTN2 PLTVDFEEFGWDWIIAPKRYRANYCSGECEFLHPQQYPHAHLVNQANPRGTAGPRCTPTK 328

S.aurataMSTNb PLTVDFEEFGWDWIIAPKRYRANYCSGECEFMHLQQYPHAHLVNKANPRGTAGPCCTPTK 328

D.rerioMSTN2 PLTVDFEDFGWEWITAPKRYKANYCSGD----FLQKYPHSHIVNKANPIFFAGPCCILTK 335

** ***. ***:** **: *:* **:*: :. *:***:*:*::.. .** * **

B.belcheriGDF8/11 MSNISMLYFDDDGNIIYAKLPDMKVDRCGCS 419

B.floridae GDF8/11 MSNISMLYFDDDGNIIYAKLPDMKVDRCGCS 419

M.musculusMSTN MSPINMLYFNGKEQIIYGKIPAMVVDRCGCS 376

R.norvegicusMSTN MSPINMLYFNGKEQIIYGKIPAMVVDRCGCS 376

H.sapiensMSTN MSPINMLYFNGKEQIIYGKIPAMVVDRCGCS 375

G.gallusMSTN MSPINMLYFNGKEQIIYGKIPAMVVDRCGCS 375

M.gallopavoMSTN MSPINMLYFNGKEQIIYGKIPAMVVDRCGCS 362

R.norvegicusGDF11 MSPINMLYFNDKQQIIYGKIPGMVVDRCGCS 405

M.musculusGDF11 MSPINMLYFNDKQQIIYGKIPGMVVDRCGCS 405

H.sapiensGDF11 MSPINMLYFNDKQQIIYGKIPGMVVDRCGCS 407

D.rerioGDF11 MSPINMLYFNDKQQIIHGKIPGMVVDRCGCS 390

T.rubripesMSTN1 MSPINMLYFNQEQQIIYGKIPSMVVDRCGCL 376

S.aurataMSTN MSPINMLYFNRKEQIIYGKIPSMVVDRCGCS 385

D.rerioMSTN1 MSPINMLYFNGKEQIIYGKIPSMVVDRCGCS 374

T.rubripesMSTN2 MSPINMLYFNRKEQIIYGKIPSMVVDHCGCS 359

S.aurataMSTNb MSPINMLYFNRKEQIIYGKIPSMVVDHCGCS 359

D.rerioMSTN2 MSPINMLYFNDREQIIYGKIPSMVVDLCGCS 366

** *.****: :**:.*:* * ** ***

Fig.S1.Comparison of deduced amino acid sequences of GDF8/11 of amphioxus with GDF8/MSTNs and GDF11s of other organisms. *Represents identical amino acid. H. sapiens MSTN (NP_005250), M. musculus MSTN (NP_034964), R. norvegicus MSTN (AAB86691), G. gallus MSTN (AAR18244), M. gallopavoMSTN (AAB86692), S. aurata MSTN (AAK53545), S. aurata MSTNb (AAL05943), T. rubripes MSTN1 (AAR88255), T. rubripes MSTN2 (AAR88254), D.rerio MSTN1 (AAB86693), D. rerio MSTN2 (AY693972), H. sapiensGDF11 (NP_005802), M. musculus GDF11 (NP_034402), R. norvegicus GDF11 (XP_343149), D. rerio GDF11 (AAN03678).The nine conserved Cystein residues are shaded.