Supplementary Material
Table 1. Primers used in the quantitative RT-PCR.
Amplicon
Primer
Sequence
FBXO31_1F
GTGAGCTCCAGGAATGAGGA
FBXO31_1R
GATTTCAGCTCCAGCCAGAC
FBXO31_2F
GTTCTCCACCAAGTGCAACC
FBXO31_2R
ATTGAAGTTGCGCTGGTTCT
FBXO31_3F
GCCGTGAGGAGTATGGTGTT
FBXO31_3R
CGTTCAGCAGTCCTCCGTAT
1
2
3
Table 2. Primers used for validation of exon 7 mutation:
Primer
Sequence
FBXO31_7F
GGCCCTGGTTGAGCTGAG
FBXO31_7R
GGATCAGTGTCTGCCCAAGT
Table 3. Whole exome sequencing statistics from individual IV:8. The single SNP remaining
after filtering out dbSNP131 known SNPs, non-coding variants, heterozygous variants, and
finally variants not within the two HBD regions on 16q, was in the gene KATNB1, c.599A>C;
p.Asn200Thr, had low coverage (N=3 reads), and is predicted by PolyPhen2, ConDel, and others
to be neutral .
WES coverage
Number of all variations
Number of Homozygous variations
Mean coverage
Median coverage
% variants with ≥3X coverage
% variants with ≥10X coverage
WES variants
Number of SNPs
Number of Indels
Number of homozygous SNPs
Number of homozygous Indels
Number of homozygous coding SNPs
Number of homozygous coding Indels
Number of homozygous coding SNPs in HBD regions
Number of homozygous coding Indels in HBD regions
Number of coding exons with less than 3X coverage in HBD regions
WES variants filtered for SNPs in dbSNP131
Number of homozygous coding SNPs
Number of homozygous coding Indels
Number of homozygous coding SNPs in HBD regions
Number of homozygous coding Indels in HBD regions
FBXO31 protein sequence:
Wild-type FBXO31 protein
MAVCARLCGVGPSRGCRRRQQRRGPAETAAADSEPDTDPEEERIEASAGV
GGGLCAGPSPPPPRCSLLELPPELLVEIFASLPGTDLPSLAQVCTKFRRI
LHTDTIWRRRCREEYGVCENLRKLEITGVSCRDVYAKLLHRYRHILGLWQ
PDIGPYGGLLNVVVDGLFIIGWMYLPPHDPHVDDPMRFKPLFRIHLMERK
AATVECMYGHKGPHHGHIQIVKKDEFSTKCNQTDHHRMSGGRQEEFRTWL
REEWGRTLEDIFHEHMQELILMKFIYTSQYDNCLTYRRIYLPPSRPDDLI
KPGLFKGTYGSHGLEIVMLSFHGRRARGTKITGDPNIPAGQQTVEIDLRH
RIQLPDLENQRNFNELSRIVLEVRERVRQEQQEGGHEAGEGRGRQGPRES
43657
22136
13.10X
9X
91.8
46.5
41769
1888
21094
1042
20042
999
369
10
211
298
13
1
0
QPSPAQPRAEAPSKGPDGTPGEDGGEPGDAVAAAEQPAQCGQGQPFVLPV
GVSSRNEDYPRTCRMCFYGTGLIAGHGFTSPERTPGVFILFDEDRFGFVW
LELKSFSLYSRVQATFRNADAPSPQAFDEMLKNIQSLTS
The epitope sequence where the C-terminal anti-FBXO31 antibodies (Bethyl Laboratories and
Novus Biologicals) bind is indicated in blue (residues 489 to 539).
Mutated Protein
MAVCARLCGVGPSRGCRRRQQRRGPAETAAADSEPDTDPEEERIEASAGV
GGGLCAGPSPPPPRCSLLELPPELLVEIFASLPGTDLPSLAQVCTKFRRI
LHTDTIWRRRCREEYGVCENLRKLEITGVSCRDVYAKLLHRYRHILGLWQ
PDIGPYGGLLNVVVDGLFIIGWMYLPPHDPHVDDPMRFKPLFRIHLMERK
AATVECMYGHKGPHHGHIQIVKKDEFSTKCNQTDHHRMSGGRQEEFRTWL
REEWGRTLEDIFHEHMQELILMKFIYTSQYDNNLPPHLPAAQPPRRPHQA
WPLQRYLWQPRPGDCDAQLPRPACQGHQDHGRPQHPRWAADSGDRPEASD
PAARPREPAQLQ*
Amino acid sequence after the indel mutation and frameshift is indicated in red