supplementary information - nature · dcn ccttctggcacaagtctcttgg tcgaagatgacactggcatcgg ......
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W W W. N A T U R E . C O M / N A T U R E | 1
SUPPLEMENTARY INFORMATIONdoi:10.1038/nature22978
Supplementary Table 1 List of primers used for mice real-time PCR experiments
Gene Forward primer Reverse primer
Agrn TTCGATGGTCCTTGTGACCC AGATAGGTGTGTGTTGGGCG
Tgfbi CATCGACGCCCAGATGAAGA TGGTGAACAGGGTCCCAAAC
Vcan CCAGTGTGAACTTGATTTTGATGAA AACATAACTTGGGAGACAGAGACATCT
Col18 GTGCCCATCGTCAACCTGAA AGTTGACCCTGGGAGCCAGA
Dcn CCTTCTGGCACAAGTCTCTTGG TCGAAGATGACACTGGCATCGG
Lum TCGAGCTTGATCTCTCCTAT TGGTCCCAGGTCTTACAGAA
Hprt TGGCCGGCAGCGTTTCTGAG GTCGGCTCGCGGCAAAAAGC
Acta1 GACATCAAAGAGAAGCTGTG ACTCCATACCGATAAAGGAAG
Dag1 CGGAGGAGCGAACACCTG GTTGGATCCTCACCCTCTGC
CD90 GTCAGGCTGGTCACCTTCTG AACTCTTGGCACCATGAACC
Myh6 GCTGGGCTCCCTGGACATTGAC CCTGGGCCTGGATTCTGGTGAT
Pecam-1 CCAGAAACATCATCATAACCG CATCGCCACCTTAATAGTTG
SUPPLEMENTARY INFORMATION
2 | W W W. N A T U R E . C O M / N A T U R E
RESEARCH
Supplementary Table 2 List of primers used for human real-time PCR experiments
Gene Forward primer Reverse primer
GAPDH GGAGCGAGATCCCTCCAAAAT GCAAATGAGCCCCAGCCTTC
CACNA1C AATCGCCTATGGACTCCTCTT GCACTAAAAAGCCCCACAACC
KCNJ2 CTGGGTCTTGGGAATTCTGGTT GAGAACATGTCCTGTTGCTGGC
GJA1 TCATTAGGGGGAAGGCGTGA CTGGGCACCACTCTTTTGCTT
SCN5A TCTCTATGGCAATCCACCCCA TGAAGAGCGAGTGAACCAGA
S100A1 TTCCTGGATGCCCAGAAGGATG CCGTCTCCATTCTCGTCTAGC
MYOM2 GCTTTTGCAGAGAAGAATCGTG CATGCTGACGTACTTGGCCT
MYOM3 TGAAGCCAGTCTTTGCTCGT ACCTCAGTAGGCTCCCATCT
COX6A2 ACTCCTATCTCCACTCGGGC GGTAGGGCTTGGTGCGGAT
CASQ2 TTGCCATCCCCAACAAACCT AGAGTGGGTCTTTGGTGTTCC
MYL2 TTGGGCGAGTGAACGTGAAAA TCCGCTCCCTTAAGTTTCTCC
MYL7 GCCCAACGTGGTTCTTCCAA GCTGAAGGCTTCTTTGAACTCC
TNNI3 CCTCACTGACCCTCCAAACG GAGGTTCCCTAGCCGCATC
TNNI1 ATGCCGGAAGTCGAGAGAAAA TCGTATCGCTCCTCATCCAC
Supplementary Table 3 List of primers used for mouse genotyping
Gene Forward primer Reverse primer
Cre GGCCAGCTAAACATGCTTCA ACACCAGAGACGGAAATCCATC
Rosa26-tdTomato GGCATTAAAGCAGCGTATCC CTGTTCCTGTACGGCATGG
Agrinflox(E6-NEO) CGGACACACATATGCTAGTG CCGCCTCAGGACTCTTCCTTT
Agrinwt(E6-E7) CGGACACACATATGCTAGTG CACTGAGGTAGGTGACGCCA
W W W. N A T U R E . C O M / N A T U R E | 3
SUPPLEMENTARY INFORMATION RESEARCH
Agrin
GAPDH
Agrin
Tubulin
Tnni3
Tnni1
Tubulin
Myh6
Myh7
Tubulin
WT Agrin-cKO
WT Agrin-cKO
Fig 1j Fig 2d
Fig 4a
Extended Data Fig. 5a Extended Data Fig. 7a
Fig 4d
Fig 4e
Fig 4h
Fig 4i Fig 4k
Fig 4l
Fig 2fP1 P8 12W
Dag1
GAPDH
MuSK
Tubulin
IB: α-dystroglycan IB: β-dystroglycanIB: rec-Agrin
IB: Desmin
Myofibrils
Cyt
osol
IB: Myh6IB: Actn2
IB: SyntrophinIB: Dystrophin
pERK
gERK
pERK
gERK
P1 P8
P1PC P8
WT Agrin-cKO
Agrin
Tubulin
Time (hrs)
T0
T2
T48
0 2 242 48
0 242 48
0 2 240 2 48
Fractions23211917152321191715
2321191715
2321191715
2321191715
0
- + + + + +AgrinTime 5 15 30 60 120
Dag1 Ab
Agrin -
- -
- +
+ +
+
Dystrophin
Yap
Yap
β-Dystroglycan
β-Dystroglycan
Syntrophin
Syntrophin
α-Dystroglycan
rec-Agrin
rec-Agrin
IP AB- +
IP AB- +
IP ABAgrin
-- 0
+ + + +2 24 48
75
75
100
135
100
135
180245245
245245
180245245
63
48
6348
3563
63
48
35
48
35
48
48
63
63
135
10075
135
10075
135
100
75
100
135
48
63
48
63
48
63
180
245245
180
135
135
100
75
135
100
75
135
100
75
135
100
75135
245
48
100
75
245
48
48
135
100
75
75
63
63
245
180135
48
3525
245180135
100
48
35
25
460
238
171
268
460
238
171
71
55
41
268
71
55
41
31
55
41
31
71
55
41
31
0--
1 6 24 48 72 96Time postMI (Hours)
Agrin + + + + + + +
24518013510075
63
48
35
245180135100
75
63
48
180
135
100
245
180
135100
75
75
63
48