Supplemental Table 1. Microsatellite primers. Physical positions are from Ensembl Release 38 (Ensembl 2006).

Primer pair name / Position (Mb) / Forward primer / Reverse primer
D1Mit265 / 148.77 / AAATTTGGCTTGATATAAAGCAGC / CCAATGTGAGGCATATTGTCA
D1Mit200 / 150.24 / GCCATGTTCATGTACATAGGTAGG / ATGGATGGATGGTTTTCCTG
D1Mit513 / 151.82 / GAGACCCTTGAAATTCACAACC / GTGTTTGATAGCTTTGATGAGGG
D1Mit396 / 153.08 / GGAGAAATTCATATGACTGATATTTTG / GGTTCAGTTTGATAACTTATACCAACA
D1Mit42 / 154.18 / CTCAGGCACCATTCTAAACATG / ATAGGGCAAAAAACATTCTTGC
D1Icp156 / 154.99 / CCAGAGATCAGCCAAGAAGA / TCAGGGAGAGAGTACGGATG
D1Icp157 / 155.04 / CTCCCAACAGATTCCACCTT / ATTCTGTGGCTCTTGTGGTC
D1Mit451 / 155.92 / TCAACTAATTCATTTTTTAAAGGGTAC / TGCATGAACTTTTACATGTGTACA
D1Mit202 / 157.00 / CCATAAGCCTCCTCTTTCCC / AAAATGAACTCAGCGGGTTG
D1Mit290 / 157.92 / GTCAACTTCTGATCTCCTCATGC / CATCTATGCATGTGTGATGTGG
D1Icp5 / 158.09 / TTTTGTTTCTGCAATTTTGTT / TCATCCATTCAGTTTCATAGGT
D1Icp6 / 158.11 / AAGCCTCATTCAAGGTTTCC / CTCAGAACCTCCTCCTTCGT
D1Icp7 / 158.18 / AAATGTATGTTCGATTGTAGGG / TCTTGGAAAAGAAACTCAGAAA
D1Icp1 / 158.19 / GACAGGAGTGGCTGATGATG / GGGCCTGGCTAAAGTATGTC
D1Icp4 / 158.22 / CATGTGCAAAATGGTAGGTC / CAGAGCTTGTCACTTTGTGG
D1Mit227 / 158.23 / TAACAAGTGAAGTCCATGGGC / AGGGGAGACAGGGACAGG
D1Icp8 / 159.03 / AATATCACTGCTGCCCAAAG / TCTTGCTGCCCATTAAACTC
D1Icp9 / 159.06 / TTCCAAATCCTGCCACTATC / CAAACATCCAGTCCTTCTCC
D1Icp10 / 159.12 / TTGATTTTCGGACTCCTATGT / GCAAAACTTCTGTGGAAGGA
D1Icp11 / 159.32 / TGTCACTGAAAACATTTCCTAA / GCCAAATATAAGCTCCCAAG
D1Mit219 / 159.52 / AAGGATGCAGTCTTTTAAATAATGG / GCCAAATAAGCCTAAAGGTTTT


Supplemental Table 2. Primers used to sequence PAPPA2 coding region. All primers were designed to span introns and were used to sequence cDNA, except Cr02 and Cr01, which were designed for direct sequencing of genomic DNA.

Primer pair name / Forward primer / Reverse primer / Product size (bp)
Cr02 / acacagcagcaaaccctgt / GGACTGTCACTATCTCCATGCT / 452
Cr21 / TGTCTCCACCATTGCTACCT / CATCTCTCCACCCCTTGTC / 500
Cr22 / CAGAGGGAGGACAGAGCAA / GTAAAGGTGACAGAATCTCAGG / 600
Cr23 / GGGCTCCATTTAGAGATGAC / ACTCATGTATGCCCTCTTGG / 605
Cr24 / AAGTGACTGATGTCCGAAAGA / GTGTCATTAGCTGGCTCTGG / 433
Cr25 / TGACACTGCTCCCACACC / GGAGTCCAGTAACCTGAAGAG / 417
Cr26 / CCAATCAGTGGAGTTGTGTATG / CATGTGGAGTTTGATGGTGA / 441
Cr27 / CCTCTCCATGAACTCTTCCA / TCTACCTTCCCATCTCCACA / 447
Cr28 / GCTGAAGCTGTAGGGGAAC / CTGAAGACACAGGGAAACCA / 446
Cr29 / GGTAACAGGAGAACCTCATTCA / TACTGGGAGCTGAAGAACTGG / 482
Cr11 / GGAACCTATGGACTGTCGTG / AATTCACCAAGGAGGAGTCTG / 478
Cr12 / TTCTCCTTACTTGTTCCTCTGG / GTTCCCAGCTTCCATCATC / 429
Cr13 / CTGCTCCTGCCACATTTC / ACTTTGACAGGTTCCATCCA / 521
Cr14 / AAGTGTGGACAGGGATATGG / GACAGGTGCATTCATCCAAG / 352
Cr01 / gaggaaaattaaagccagttga / actgacagcgtgcagagt / 474

6

Supplemental Table 3. Primer and probe sequences for quantitative PCR. TaqMan MGB probes (Applied Biosystems) labelled at the 5' end with FAM were used.

Gene / Forward primer / Reverse primer / Probe
PAPPA2
(probe a) / CAGCCCTTGTCATTGACTGTGT / AGCAACCCAGCCATCATTG / AGCAGTTATCAAACACACCT
PAPPA2
(probe b) / TACACATGATGGGGAGCACACT / GTGTGAAAACCTGCAAGGAGAAT / ACCGATCCCATATCC
IGFBP-5 / AAGAGCTACGGCGAGCAAAC / GTGGTGGGTTCCTCGTGTTC / CGAGAGTCTCTCTCTATCTT
b-actin / CCTAAGGCCAACCGTGAAAAG / CTACGTACATGGCTGGGGTGTT / ACCCAGATCATGTTTGAGAC

6

Supplemental Table 4. Single nucleotide polymorphisms (SNPs) in the coding region of PAPPA2.

5' flanking sequence / Codon polymorphism
C57 allele/ DBA allele / 3' flanking sequence / Amino acid change
C57 / DBA
CCCACAGCAGAGACCCAGAGG / AAG/AGG / GGCCAGTCCAAGTCTAGGCTT / Lys / Arg
CGGAGAGTAGAGACCTTTAAC / TCC/TAC / CAAGGAGGATTGCCTGTACTG / Ser / Tyr
GTGCTGGCTGAGATTCCCCGG / GAG/AAG / GCGTTCACAGTGGAAGCCTGG / Glu / Lys
AATGTCTACTTCGCCAGCTCA / GTC/GTT / CGGGAAGACCTTGCAGGTTCT / Synonymous
TTGCTTTTTCAATACCCGGTC / CAG/CAA / GCGGATACCCTCACCCTCTGG / Synonymous
CCCATGGTGGTGACACATCCA / CAC/CAT / AGAAAGTTCACAGATACGGAG / Synonymous
TTCACAGATACGGAGGTCATC / CCT/CCC / GGGCAGATGTATCAGTACCAA / Synonymous
GATACGGAGGTCATCCCTGGG / CAG/AAG / ATGTATCAGTACCAAGTTCAA / Gln / Lys
TACCAAGTTCAAGCTGAAGCT / GTA/GGA / GGGGAACTCGGAGAAGCTTCA / Val / Gly
GGAGAAGCTTCACCTCCTCTG / AGC/AGG / CACATTCACGGAGGCCCTTAC / Ser / Arg
CCTCCTCTGAGCCACATTCAC / GGA/GGG / GGCCCTTACTGTGGAGATGGG / Synonymous
GATGGGAAGGTAGAAAGGAGT / ATC/ATT / GGAGAGATGTGTGATGATGGA / Synonymous
GAGGAAGGCTTCAATTGTGCA / GGG/GAG / GAGCCAAGCCTTTGCTACAGG / Gly / Glu
AAATGTCCTGTTTCCCTGGTA / ACA/ATA / GGAGAACCTCATTCAATGATC / Thr / Ile
TCAATGATCTGTACTTCATGG / CAC/CGC / CCGGATTCATCCCCATATCAT / His / Arg
GGGGTTGCAGTGGCAATTTAC / ATT/GTT / TTCTTGGCATCGGATGGCGTG / Ile / Val
CTGGTTATCAATGTGAGCCAT / CAT/CCT / GTGAATGTCTACCCCCAACAT / His / Pro
TGTGGAGAACAGGGAAGATGT / GCG/GTG / CCATTGCTACTCGATCATACG / Ala / Val
GCGCCATTGCTACTCGATCAT / ACG/ACA / GACATGGTGAATTGTACCTCC / Synonymous
GACATGGTGAATTGTACCTCC / AGT/AGC / AGCCCAGGTCACATGGAGTGC / Synonymous
ACAGACTTTCTGAAACAATGT / TCC/TCT / ATCTCGTGTGTCCCACCGGCT / Synonymous
TGTTCCATCTCGTGTGTCCCA / CCG/CCA / GCTAAGCTCCAAGAGTCAGGA / Synonymous
AAGCTCCAAGAGTCAGGACTG / AAC/AAT / CCATGGCTGACATGTCTTGAA / Synonymous
GAGACTTCAGGAAGTCAAGGC / AAG/AAA / AACAAGTTCCTGAAGATCCAG / Synonymous
AAGTTCCTGAAGATCCAGTGC / CTG/CTA / GATGATGGAAGCTGGGAACAA / Synonymous
AGCTGGGAACAAGGGAGCTGC / GTC/ATC / CCCGTGGTTTGTGAGCCTCCT / Val / Ile
CCTAGTGACCCTGTGATACTA / CCA/CCG / GAGAATGTGACTACTGACACT / Synonymous
TGTACCGGCAGGCGCCAGTGG / CAC/TAC / CCAGATCCCTCTCTGGTGCAC / His / Tyr
GACTGCTGCTCTTCCACACTC / TCT/TCG / TCCAAGAAGGTCATTCCATTT / Synonymous

6