Identification of Zebrafish Magnetoreceptor and Cryptochrome Homologs

SCIENCE CHINA Life Sciences Supporting Information

Identification of zebrafish magnetoreceptor and cryptochrome homologs

This file includes: Figures. S1 to S4

Table. S1

M-SASIARATVRAVSRRKILSTRAALTLTPSAVNKVKQLLDNKPEYIGLKVGVRTRGCNGLTYTLDYTKNKEQSDEEVLQDGVR M-SASVVRATIRAVSKRKLQPTRAALTLTPAAVNKVKKLLQDKPDNIGLKVAVRTRGCNGLSYYLEYAKSKGPSDEEVVQDGVR Y------GIP------CLPLTPSAVHKIKQLLKDKPEHVGVKVGVRTRGCNGLSYTLEYAKTKGDSDEEVAQDGVR M-ASSVVRATVRAVSKRKIQATRAALTLTPSAVQKIKQLLKDQPEHVGVKVGVRTRGCNGLSYTLEYTKSKGDSDEEVVQDGVR M-SASLVRATVRAVSKRKLQPTRAALTLTPSAVNKIKQLLKDKPEHVGVKVGVRTRGCNGLSYTLEYTKTKGDSDEEVIQDGVR MATRVVATATVRAVKGRKLIPTRAALTLTPAAVLRIKTLLQDKPDMVGLKVGVRQRGCNGLSYTLDYASQKDKLDEEVVQDGVK species % hMAGR % fMagR VFIEKKAQLTLLGTEMDFVETKLSSEFVFNNPNIKGTCGCGESFNI zebrafish 84 74 VFIEKKAQLTLLGTEMDYVEDKLSSEFVFNNPNIKGTCGCGESFNI frog 86 78 VFIEKKAQLTLLGSEMDYVEDKLSSEFVFNNPNIKGTCGCGESFNI turtle 75 75 VFIEKKAQLTLLGTEMDYVEDKLSSEFVFNNPNIKGTCGCGESFNI chicken 91 75 VFIEKKAQLTLLGTEMDYVEDKLSSEFVFNNPNIKGTCGCGESFNI human 100 78 VFIDKKAQLSLLGTEMDFVESKLSSEFVFNNPNIKGTCGCGESFSM fruitfly 74 100 Supplementary Figure 1. Sequence alignment of MagR proteins. % identity values are from comparisons between the human MAGR (hMAGR) or fruitfly MagR (fMagR) and their counterparts from other organisms. For protein accession numbers see Figure 1. 2

1 46 58 76 82 96 107 128 M--VHWFRKGLRLHDN--PSLRDSI--CVYILDPWFAGS--SNVGIS--RWRFLLQCLEDLDASLRKLN---SRLFVIRGQPTDVFPRLFKEWNIN- RLSYEYDSEPFGKERDAAIKKLANEAGVE M--VHWFRKGLRLHDN--PSLRDSI--CVYILDPWFAGS--SNVGIS--RWRFLLQCLEDLDASLRKLN---SRLFVIRGQPTDVFPRLFKEWK- ISRLSYEYDSEPFGKDRDAAIRKLATEAGVE M--IHWFRKGLRLHDN--PALQEAV--CVYFLDPWFAGS--SNLGVN--RWRFLLQCLDDLDSNLRKLN---SRLFVVRGQPANVFPRLFKEWK- ISRLTFEYDSEPFGKERDAAIKKLAMEAGVE M--VHWFRKGLRLHDN--PALLEAL--CVYFLDPWFAGA--SNLGVN--RWRFLLQSLEDLDASLRKLN---SCLFVIRGQPADIFPRLFKEWKVS- RLTFEFDSEPFGKERDAAIKKLACEAGVE M--VHWFRKGLRLHDN--PALQEAL--CVYILDPWFAGS--ANVGVN--RWRFLLESLEDLDTSLRKLN---SRLFVVRGQPTDVFPRLFKEWN- VTRLTFEYDSEPYGKERDAAIIKMAQEYGVE M--IHLFRKGLRLHDN--PSLLGAL---VYVLDRVFLQG---AMHMGALRWRFLLQSLEDLDTRLQAIG---SRLFVLCGSTANILREL- VAQWGITQISYDTEVEPYYTRMDKDIQTVAQENGLQ M--IHWFRKGLRLHDN--PALIAAL---LFLLDPWFPKN--TRIGIN--RWRFLIEALKDLDSSLKKLN---SRLFVVRGSPTEVLPKLFKQWK- ITRLTFEVDTEPYSQSRDKEVMKLAKEYGVE M--VHWFRKGLRLHDN--PALRECI--CVYILDPWFAGS--SNVGIN--RWRFLLQCLEDLDANLRKLN---SRLFVIRGQPADVFPRLFKEWK- ITKLSIEYDSEPFGKERDAAIKKLASEAGVE M--VHWFRKGLRLHDN--PALLAAL--CVYILDPWFAAS--SSGGVN--RWRFLLQSLEDLDSSLRKLN---SRLFVVRGQPADVFPKLFKEWGVS- RLTFEYDSEPFGKERDAVIMKLAKEAGVE M--KRWISSKMKLPAE--TALQASL—-VVMYRGAWLQVS--INVGLFFLLHRFLLQCLEDLDANLRKLN---SRLFVIRGQPADVFPRLFKEWNI- AKLSIEYDSEPFGKERDAAIKKLASEAGVE S------LSYTQT--TAADCLF------HHRFLLQSLEDLDNSLRKLG---SCLFVVRGQPTDVF- PRLFKEWGVTRLTFEYDSEPFGKERDAAIIKLAKEAGVE M--VHWFRKGLRLHDN--PALRECI--CVYILDPWFAGS--SNVGIN--RWRFLLQCLEDLDANLRKLN---SRLFVIRGQPADVFPRLFKEWSI- AKLSIEYDSEPFGKERDAAIKKLASEAGVE M--VHWFRRGLRLHDN--PALQAAL--CIYILDPWFAAS--SAVGIN--RWRFLLQSLEDLDNSLRKLN---SRLFVVRGQPTDVF- PRLFKEWGVTRLTFEYDSEPFGKERDAAIIKLAKEAGVE M--VHWFRKGLRLHDN--PALKECI--CVYILDPWFAGS--SNVGIN--RWRFLLQCLEDLDANLRKLN---SRLFVIRGQPADVF- PRLFKEWNITKLSIEYDSEPFGKERDAAIKKLATEAGVE M--VHWFRKGLRLHDN--PALLAAV--CVYILDPWFAAS--SSVGIN--RWRFLLQSLEDLDTSLRKLN---SRLFVVRGQPADVF- PRLFKEWGVTRLTFEYDSEPFGKERDAAIMKMAKEAGVE M--VIWFRHGLRLHDN--PALLAAL--VVFIFDGESAGT--KNVGYN--RMRFLLDSLQDIDDQLQAATDGRGRLLVFEGEPAYIFRRLHE- QVRLHRICIEQDCEPIWNERDESIRSLCRELNID

VIVRISHTLYDLDKIIELNGGQSPLTYKRFQTLISRMEAVETPAETITAE--VMGPCTTPLSDDHDEKFGVPSLEELGFDTEGL--- SSAVWPGGETEALTRLERHLER---KAWVANFERPRMN VFVRISHTLYDLDKIIEFNGGQSPLTYKRFQTLISRMDPVEMPAETITAE--IMGKCSTPVSDDHDDKFGVPSLEELGFETEGL--- STAVWPGGETEALTRLERHLER---KAWVANFERPRMN VIVKTSHTLYNLDKIIELNGGQPPLTYKRFQTLISRMDPPEMPVETLSNS--IMGCCVTPVSEDHGDKYGVPSLEELGFDIEGL--- PSAVWPGGETEALTRIERHLER---KAWVANFERPRMN VIVKISHTLYDLDRIIELNGGQSPLTYKRFQTLVSSMEPPDPPLASPDRG--MMGKCVTPISENHRDKYGVPLLEELGFDTEGL---APAVWPGGE- SEALKRMERHLGPDSTVAWQENFERPKMN TVVRNTHTLYNPDRIIEMNNHSPPLTFKRFQAIVNRLELPRKPLPTITQE--QMARCRTQISDNHDEHYGVPSLEELGFRTQGD--- SLHVWKGGETEALERLNKHLDR---KAWVANFERPRIS TYTCVSHTLYDVKRIVKANGGSPPLTYKKFLHVLSVLGEPEKPARDVSIE--DFQRCVTPVDVDR--VYAVPSLAHLGLQVEA-----EVLWPGGE- SHALQRLEKHFQS---QGWVANFSKPRTI VTPKISHTLYNIDRIIDENNGKTPMTYIRLQSVVKAMGHPKKPIPAPTNE--DMRGVSTPLSDDHEEKFGIPTLEDLGLDTSSL---GPHLFPGGE- QEALRRLDEHMER---TNWVCKFEKPKTS VIVRISHTLYDLDKIIELNGGQPPLTYKRFQTLISKMDPLEIPVETITAE--VMEKCTTPVSDDHDEKYGVPSLEELGFDTEGL--- PSAVWPGGETEALTRLERHLER---KAWVANFERPRMN VIVENSHTLYDSDRVIELNGHSPPLTYKRFQAIISRMELPRRLAPSVTRQ--QMEACRAEIKRNHDETYGVPSLEELGFHSENK---GPAI- WPGGETEALARLDRHLER---KAWVANYERPRMS VIVRISHTLYDLDKIIELNGGQPPLTYKRFQTLISRMEPLEMPVETITAE--VMEKCTTPVSDDHDEKYGVPSLEELGFDTDGL--- PSAVWPGGETEALTRLERHLER---KAWVANFERPRMN VVIENSHTLYDLDRIIELNGHKPPLTYKRFQTIISRMELPKKPVSSITSQ--QMEKCKSEIQDNHDDTYGVPSLEELGFPTDGL--- APAVWKGGETEALARLDKHLER---KAWVANYERPRMN VIVRISHTLYDLDKIIELNGGQPPLTYKRFQTLISRMEPLEMPVETITPE--VMQKCTTPVSDDHDEKYGVPSLEELGFDTDGL--- PSAVWPGGETEALTRLERHLER---KAWVANFERPRMN VVIENSHTLYDLDRIIELNGNKPPLTYKRFQAIISRMELPKKPVSSIVSQ--QMETCKVDIQENHDDVYGVPSLEELGFPTDGL--- APAVWQGGETEALARLDKHLER---KAWVANYERPRMN VIVRISHTLYDLDKIIELNGGQPPLTYKRFQTLISKMEPLEIPVETITSE--VIEKCTTPLSDDHDEKYGVPSLEELGFDTDGL--- SSAVWPGGETEALTRLERHLER---KAWVANFERPRMN VVTENSHTLYDLDRIIELNGQKPPLTYKRFQAIISRMELPKKPVGLVTSQ--QMESCRAEIQENHDETYGVPSLEELGFPTEGL--- GPAVWQGGETEALARLDKHLER---KAWVANYERPRMN FVEKVSHTLWDPQLVIETNGGIPPLTYQMFLHTVQIIGLPPRPTADARLEDATFVELDPEFCRSLKLFEQLPTPEHFNVYGDNMGFLAKIN- WRGGETQALLLLDERLKVEQ-HAFERGFYLPNQA

ANSLLASPTGLSPYLRFGCLSCRLFYFKLTDLYRKVKKNSSPP------LSLYGQLLWREFFYTAATNNPRFDKMEGNPICVQIPWDK-NPEALAK- WAEGRTGFPWIDAIMTQLRQEGWIHHLA ANSLLASPTGLSPYLRFGCLSCRLFYFKLTDLYRKVKKNSTPS------LSLYGQLLWREFFYTAATNNPHFDKMEFNPICVQIPWDR-NPEALAK- WAEGQTGFPWIDAIMTQLRQEGWIHHLA ANSLLASPTGLSPYLRFGCLSCRLFYFKLTDLYRKVKKTSTPP------LSLYGQLLWREFFYTAATTNPRFDKMEGNPICVRIPWDK-NPEALAK- WAEAKTGFPWIDAIMTQLRQEGWIHHLA ASPLMASPLGLSPYLRFGCLSCRLFYCKLTQLYKKVKKNMNPS------ISLYDKILWREFFYTAATNNPRFDRMEGNPICIRIPWDR-NAEALAK- WAEAKTGFPWIDAIMMQLRQEGWIHHLA GQSLFPSPTGLSPYLRFGCLSCRVFYYNLRDLFMKLRRRSSPP------LSLFGQLLWREFFYTAGTNNPNFDHMEGNPICVQIPWDH-NPEALAK- WAEGRTGFPWIDAIMTQLRQEGWIHHLA PNSLLPSTTGLSPYLSLGCLSVRTFYHRLNCIYAQSKNHSLPP------VSLQGQVLWREFFYTVASATPNFTKMEGNSICLQIDWYH-DPERLEK- WRTAQTGFPWIDAIMTQLLQEGWIHHLA PNSLIPSTTVLSPYVRFGCLSARTFWWRLADVYRG-KTHSDPP------VSLHGQLLWREFFYTTAVGIPNFNKMEGNSACVQVDWDN-N- PEHLAAWREARTGFPFIDTIMTQLRQEGWIHHLA ANSLLASTTGLSPYLRFGCLSCRLFYFKLTDLYKKVKKNSSPP------LSLYGQLLWREFFYTAATNNPRFDKMDGNPICVQIPWDR-NPEALAK- WAEGRTGFPWIDAIMTQLRQEGWIHHLA ANSLLASPTGLSPYLRFGCLSCRLFYYRLQELYQKVKKNSPPP------LSLYGQLLWREFFYTAATNNPKFDQMEGNPICVQIP- WDK-NPKALAKWTEGKTGFPWIDAIMTQLRQEGWIHHLA ANSLLASPTGLSPYLRFGCLSCRLFYFKLTDLYKKVKKNSSPP------LSLYGQLLWREFFYTAATNNPRFDKMEGNPICVQIPWDR-NPEALAK- WAEGRTGFPWIDAIMTQLRQEGWIHHLA ANSLLASPTGLSPYLRFGCLSCRLFYYRLWELYKKVKRNNTPP------LSLYGQLLWREFFYTAATNNPKFDRMEGNPICIQIPWDR-NPEALAK- WAEGKTGFPWIDAIMTQLKQEGWIHHLA ANSLLASPTGLSPYLRFGCLSCRLFYFKLTDLYKKVKKNSSPP------LSLYGQLLWREFFYTAATNNPRFDKMEGNPICVQIPWDK-NPEALAK- WAEGRTGFPWIDAIMTQLRQEGWIHHLA ANSLLASPTGLSPYLRFGCLSCRLFYYRLWELYKKVKRNSTPP------LSLYGQLLWREFFYTAATNNPKFDRMEGNPICIQIPWDK-NPEALAK- WAEGKTGFPWIDAIMTQLRQEGWIHHLA ANSLLASPTGLSPYLRFGCLSCRLFYFKLTDLYKKVKKNSSPP------LSLYGQLLWREFFYTAATNNPRFDKMEGNPICVQIPWDK-NPEALAK- WAEGRTGFPWIDAIMTQLRQEGWIHHLA ANSLLASPTGLSPYLRFGCLSCRLFYYRLWDLYKKVKRNSTPP------LSLFGQLLWREFFYTAATNNPRFDRMEGNPICIQIPWDR-NPEALAK- WAEGKTGFPWIDAIMTQLRQEGWIHHLA LPNIHDSPKSMSAHLRFGCLSVRRFYWSVHDLFKNVQLRACVRGVQMTGGAHITGQLIWREYFYTMSVNNPNYDRMEGNDICLSIPWAKPNENL- LQSWRLGQTGFPLIDGAMRQLLAEGWLHHTL

RHAVACFLTRGDLWISWEEGMKVFEELLLDADWSVNAGSWMWLSCSSFFQQFFH--CYCPVSFGRRTDPNGDYIRRYLPVLRGFPAKYIYDPW- NAPESVQKAAKCIIGVHYPMPMVHHAEASRLN RHAVACFLTRGDLWISWEEGMKVFEELLLDADWSVNAGSWMWLSCSSFFQQFFH--CYCPVGFGRRTDPNGDYIRRYLPILRGFPAKFIYDPW- NAPESVQKVAKCIIGVHYPKPMVNHAEASRIN RHAVACFLTRGDLWISWEEGMKVFEELLLDADWSVNAGSWMWLSCSSFFQQFFH--CYCPVGFGRRTDPNGDFIRRYLPILRGFPAKYIYDPW- NAPDSVQAAAKCIIGVHYPKPMVNHAEASRLN RHAVACFLTRGDLWISWEEGMKVFEELLLDADWSVNAGSWLCHSCSSFFQQFFH--CYCPVGFGRRIDPNGDFIRRYLPVLRDFPAKYIYDPWNAPHD- VQLAAKCVIGVDYPKPMVNHAEASRLN RHAVACFLTRGDLWISWESGMKVFEELLLDADWSVNAGSWMWLSCSAFFQQFFH--CYCPVGFGRRTDPSGDYIRRYIPKLKDYPNRYIYEPW- NAPESVQKAANCIVGVDYPKPMINHAESSRLN RHAVACFLTRGDLWISWEEGMKVFEEFLLDADYSVNAGNWMWLSASAFFHKYTR--IFCPVRFGRRTDPQGEYLRKYLPVLKNFPSQYIYEPWKAPED- 3

VQLSAGCIIGKDYPRPIVSHIEASQRN RHAVACFLTRGDLWISWEEGQKVFEELLLDSDWSLNAGNWQWLSASTFFHQYFR--VYSPIAFGKKTDKHGDYIKKYLPVLKKF- PTEYIYEPWKAPRSVQERAGCIVGKDYPRPIVDHEVVHKKN RHAVACFLTRGDLWISWEEGMKVFEELLLDADWSVNAGSWMWLSCSSFFQQFFH--CYCPVGFGKRTDPNGDYIRRYLPILKGFPPKYIYDPW- NAPETVQKAAKCIIGVNYPKPMVNHAEASRLN RHAVACFLTRGDLWNSWECGVKVFDELLLDADFSVNAGSWMWLSCSAFFQQFFH--CYCPVGFGRRTDPSGDYVKRYLPVLKAFPSRYIYEPWS- APESVQKEAKCIIGIDYPKPIVNHAEASRMN RHAVACFLTRGDLWISWEEGMKVFEELLLDADWSVNAGSWMWLSCSSFFQQFFH--CYCPVGFGRRTDPNGDYIRRYLPILRGFPAKYIYDPWNAPE- SIQKVAKCVIGVNYPKPMVNHAEASRLN RHAVACFLTRGDLWISWESGVRVFDELLLDADFSVNAGSWMWLSCSAFFQQFFH--CYCPVGFGRRTDPSGDYVRRYLPKLKGFPSRYIYEPW- NAPESVQKAAKCIIGVDYPKPMVNHAETSRLN RHAVACFLTRGDLWISWEEGMKVFEELLLDADWSVNAGSWMWLSCSSFFQQFFH--CYCPVGFGRRTDPNGDYIRRYLPVLRGFPAKYIYDPW- NAPESVQKAAKCVIGVNYPKPMVNHAEASRLN RHAVACFLTRGDLWISWESGVRVFDELLLDADFSVNAGSWMWLSCSAFFQQFFH--CYCPVGFGRRTDPSGDYVKRYLPKLKGFPSRYIYEPW- NAPESVQKAAKCIIGVDYPKPMVNHAETSRLN RHAVACFLTRGDLWISWEEGMKVFEELLLDADWSINAGSWMWLSCSSFFQQFFH--CYCPVGFGRRTDPNGDYIRRYLPVLRGFPAKYIYDPW- NAPEGIQKVAKCLIGVNYPKPMVNHAEASRLN RHAVACFLTRGDLWVSWESGVRVFDELLLDADFSVNAGSWMWLSCSAFFQQFFH--CYCPVGFGRRTDPSGDYIRRYLPKLKAFPSRYIYEPWNAPE- SIQKAAKCIIGVDYPRPIVNHAETSRLN RNTVATFLTRGGLWQSWEHGLQHFLKYLLDADWSVCAGNWMWVSSSAFERLLDSSLVTCPVALAKRLDPDGTYIKQYVPELMNVPKEFVHEPWRM- SAEQQEQYECLIGVHYPERIIDLSMAVKRN 611 aa protein species % hCRY1 % hCRY2 % fCry IERMKQIYQQLSCYRGLGLLAMVPS--SS------619 Cry1aa Zebrafish 80 80 39 IERMKQIYQQLSCYRGLGLLATIPS--GI------606 Cry1ab Zebrafish 77 75 40 IERMKQIYQQLSRYRGLGLLASVPS--AEVTSVYASSGNQSSMRS659 Cry1ba Zebrafish 84 77 39 IERMRQIYQQLSRYRGLSLLATVPS--NEKRREDAEGVAFST---639 Cry1bb Zebrafish 80 71 39 IERMKQVYQQLSHYRGLSLLASVPT--KQ------598 Cry2 Zebrafish 77 79 40 LELMRQVRTEQQTTAELTRDVADDP--GEVM------558 Cry4 Zebrafish 55 56 39 ILRMKAAYAKRS------PE--KR------519 Cry5 Zebrafish 59 59 37 IERMKQIYQQLSRYRGLGLLASVPS--TH------616 Cry1 Frog 89 81 40 IERMKQTYQQLSHYRGLCILASVPS--F------570 Cry2 Frog 72 88 41 IERMKQIYQQLSRYRGLGLLATVPS--TN------666 Cry1 Turtle 83 82 39 IERMKQIYQQLSRYRGLCLLASVPS--L------538 Cry2 Turtle 82 74 39 IERMKQIYQQLSRYRGLGLLATVPS--TN------621 Cry1 Chicken 91 81 40 IERMKQIYQQLSRYRGLCLLASVPS--S------582 Cry2 Chicken 79 78 41 IERMKQIYQQLSRYRGLGLLASVPS--TN------586 CRY1 Human 100 81 40 IERMKQIYQQLSRYRGLCLLASVPS--A------614 CRY2 Human 81 100 41 MLAMKSLRNSLITP------DVVV------542 Cry Fruitfly 40 41 100

Supplementary Figure 2. Sequence alignment of Cry proteins. Length (amino acid residues, aa), protein name, species and % identity compared with human CRYs (hCRY1, hCRY2) and fruitfly Cry (fCry) are given to the end. For gene accession numbers see Figure 1.

Supplementary Figure 3. RNA expression of cry genes during embryogenesis. RNA expression of cry genes dispalyed no obvious difference at gastrula (9 hpf), pharyngula (30 hpf), and hatching (48 hpf) stages between MOmagr-injected embryos (MO) and control embryos (Cont).

Supplementary Figure 4. RNA expression of early neural maker genes. RNA expression of sox2, elavl3, and gad1b dispalyed no obvious difference at gastrula (9 hpf), pharyngula (30 hpf), and hatching (48 hpf) stages between MOmagr-injected embryos (MO) and control embryos (Cont).

Table S1 Zebrafish Genes and primers used for RT-PCR analyses Gene Accession number Primers (5’-3’) Size (bp) magr NM_001025178.2 magr-F: TCAGATGAGGAGGTGTTAC 188 magr-R: ACGGGCGTCCTGAAT cry1aa NM_001077297.2 cry1aa-F: CCGTCCAACCCTAATG 383 cry1aa-R: CTCTGCGAAGTAGTGCTG cry1ab NM_131790.4 cry1ab-F: AATCACGCTGAGGCTAGCAG 517 cry1ab-R: AAAGCGAAGCATGGAGGAGT cry1ba NM_131791.2 cry1ba-F: CAGTGGTAACAGGAGTGGGAGT 307 cry1ba-R: TTGCCGTTGTAACTTGTGC cry1bb NM_131792.2 cry1bb-F: CTGTGGGTGATGTCGGTGAA 289 cry1bb-R: AAACCTTCCATTCCTTGAAG cry2 NM_131786.2 cry2-F: AGAGTAAGACCCTCAGAGACCCC 180 cry2-R: CGTTTCTGTCGGATGGTTCGGAG cry4 XM_009295766.1 cry4-F: TACGTGCGAGGTCTGTGCTA 179 cry4-R: CCAAGATGAAAACACTCCAGATCG cry5 NM_131788.1 cry5-F: GAGCCATAACACCATTCAC 142 cry5-R: GCCGATGCGAGTGTTT 5

fgf8a NM_131281.2 fgf8a-F: TACCAGCTTTACAGCCGAACC 134 fgf8a-R: CGAACTCGACTCCCAAATG otx2 NM_131251.1 otx2-F: GCCTTGAAAATCAACTTGCCGGAGT 258 otx2-R: GATAGAGGCTGGGCTCCAAATAGAC gata4 NM_131236.2 gata4-F: TTGGACGGCTTCGCACTT 215 gata4-R: TGCCGTTCATCTTATGGTAGA gata5 NM_131235.2 gata5-F: AAGTGAACCTAGTATCGCAGC 173 gatat5-R: GAGACAGAGCACACCAGGAGT has2 NM_153650.2 has2-F: ACTTGTATGGGAAGGTTTGTAG 167 has2-R: GCAGTGAAATGCCGAACAG ntl NM_131162.1 ntl-F: TTCCGGAAGTCTTGCGTCTC 217 ntl-R: CATCGAAGAACCGCGTAGGA sall4 NM_001080609.1 sall4-F: TGTAGGTTACCATGTCGCGG 88 sal4-R: TGCAAGATCCCATTTTCTGTCG sox2 NM_213118.1 sox2-F: CGAGTCTAGTTCGAGTCCGC 193 sox2-R: GTTAATCGTCGTACCGGGCA gad1b NM_194419.1 gad1b-F: CAGGTTACCTCTTCCAGCCG 534 gad1b-R: CGTTAAACGCGTAGGCGAC elavl3 XM_017353194.1 elavl3-F: TGGCTTTGTAAACTATGTGGATCC 298 elavl3-R: CTCTGCTTCGTTCCGTTTGTCG β-actin NM_131031.1 β-actin-F: TACAGCTTCACCACCACAGC 209 β-actin-R: AGGAAGGAAGGCTGGAAGAG