SUPPLEMENTAL DATA
Supplementary Table 1: Avian ToxChip PCR Array gene list (3rd generation)
Gene
GenBank
Description
Symbol
Accession #
CYP3A37
NM_001001751
Cytochrome P450 A 37
CYP1A4
NM_205147
Cytochrome P450 1A4
UGT1A9
XM_001234353
UDP glucuronosyltransferase 1 family, polypeptide A9
SULT1B1 a
NM_204545
Sulfotransferase family, cytosolic, 1B, member 1
a
BATF3
XM_419428
Basic leucine zipper transcription factor, ATF-like 3
PDK4 a
NM_001199909
Pyruvate dehydrogenase kinase, isozyme 4
TXN
NM_205453
Thioredoxin
ACSL5
NM_001031237
Acyl-CoA synthetase long-chain family member 5
HMGCR
NM_204485
3-hydroxy-3-methylglutaryl-Coenzyme A reductase
SLCO1A2
XM_416421
Solute carrier organic anion transporter family, member
1A2
a
TTR
NM_205335
Transthyretin
DIO1
NM_001097614
Deiodinase, iodothyronine, type I
THRSP
NM_213577
Thyroid hormone responsive (SPOT14 homolog, rat)
IGF1
NM_001004384
Insulin-like growth factor 1 (somatomedin C)
NCOA3
XM_417385
Nuclear receptor coactivator 3
SULT1E1 a
XM_420616
Sulfotransferase family 1E, estrogen-preferring, member
1
CYP7B1
XM_418276
Cytochrome P450, family 7, subfamily B, polypeptide 1
CASP1 a
NM_204924
Caspase 1, apoptosis-related cysteine peptidase
(interleukin 1, beta, convertase)
CIDEC
XM_003641931
Cell death activator CIDE-3-like
HSD3B1
NM_205118
Hydroxy-delta-5-steroid dehydrogenase, 3 beta- and
steroid delta-isomerase 1
a
ALAS1
NM_001018012
Aminolevulinate, delta-, synthase 1
IL16
NM_204352
Interleukin 16 (lymphocyte chemoattractant factor)
MT4
NM_205275
Metallothionein 4
HSP90AB1 a NM_206959
Heat shock 90kDa protein 1, beta
a
CD36
NM_001030731
CD36 molecule (thrombospondin receptor)
SCD a
NM_204890
Stearoyl-CoA desaturase (delta-9-desaturase)
LBFABP a
NM_204634
Liver basic fatty acid binding protein
a
EEF1A1
NM_204157
Eukaryotic translation elongation factor 1 alpha 1
RPL4
NM_001007479
Ribosomal protein L4
a
Genes that were not included on the original ToxChip PCR Array (Porter et al. [20])
Supplementary Table 2: Raw cycle threshold (Ct) data derived from the Avian ToxChip PCR
array for 27 target genes and two control genes in hepatic tissue of 6 individuals exposed via egg
injection to the DMSO vehicle control (A) or 50 µg (116 nmol) TDCPP/g egg (B). Average Ct
values (± standard deviation) are included and blank boxes represent wells in which no
amplification was observed.
A.
Symbol
Rep 1 Rep 2
CYP3A7
27.48 23.13
CYP1A4
24.28 24.83
UGT1A9
33.02
31.1
SULT1B1
24.26 22.49
BATF3
24.49 24.05
PDK4
22.47 21.28
TXN
20 18.65
ACSL5
25.08
23.7
HMGCR
26.36 24.69
SLCO1A2
28.7 29.12
TTR
15.75 14.91
DIO1
23.18 21.97
THRSP
34.24
IGF1
34.39
33.9
NCOA3
26.74 25.99
SULT1E1
22.4 21.22
CYP7B1
25.84 24.86
CASP1
26.2 25.02
LOC100859733 24.18 22.96
HSD3B1
27.36 27.01
ALAS1
24.52 22.58
IL16
31.5 30.42
MT4
22.38
27.8
HSP90AB1
26.07
24.5
CD36
22.85 22.02
SCD
30.93 30.29
LBFABP
20.01 19.06
EEF1A1
18.78 17.59
RPL4
19.42 18.12
DMSO
Rep 3 Rep 4 Rep 5 Rep 6 AVG
SD
26.51 22.35 23.82 23.76 24.51
2.02
24.67 21.97 22.13 21.28 23.19
1.57
30.58 28.07 29.33 29.96 30.34
1.68
22.28 21.31 21.93 21.27 22.26
1.10
24.79 23.59 23.35 22.35 23.77
0.88
21.38 20.32
21.1 20.82 21.23
0.72
17.75 16.99 19.06 17.65 18.35
1.10
24.23 21.76 22.46 21.85 23.18
1.36
25.69 22.73 23.79 23.28 24.42
1.41
30.04
26.9 29.07 28.51 28.72
1.04
14.55 14.61 15.21 14.82 14.98
0.45
21.9 21.02 21.58 21.19 21.81
0.77
33.2 33.33
35 33.52 33.86
0.75
33.81 29.99 34.96 32.44 33.25
1.80
25.8
23.4 24.49 23.71 25.02
1.35
20.68 19.73 20.52 19.82 20.73
0.99
25.97 22.36 23.55 23.22 24.30
1.48
24.9 23.86
25.3 24.27 24.93
0.82
22.99 21.83 22.94 22.94 22.97
0.74
26.39 25.43 26.35 25.89 26.41
0.71
21.24 21.22 23.38 22.73 22.61
1.27
30.24 28.07 30.04 29.59 29.98
1.13
26.62 24.37 22.05 21.11 24.06
2.69
25.5 21.91
23.6 22.77 24.06
1.60
22.13
20.5 20.84 20.48 21.47
1.00
29.97 27.62 28.45 27.59 29.14
1.44
19.7
19.4 18.52 18.59 19.21
0.60
18.32 15.89 16.79 16.28 17.28
1.15
17.81 16.79 18.19 17.58 17.99
0.87
B.
Symbol
Rep 1 Rep 2
CYP3A7
22.19 20.99
CYP1A4
27.69 23.57
UGT1A9
29.69 27.13
SULT1B1
22.48 21.55
BATF3
24.54 22.76
PDK4
23.04 20.84
TXN
19.76 17.68
ACSL5
24.21 22.75
HMGCR
25.13 23.81
SLCO1A2
27.12 23.66
TTR
15.78 13.62
DIO1
22.7 20.88
THRSP
34.11
IGF1
32.97
NCOA3
25.71 24.15
SULT1E1
21.91 19.55
CYP7B1
24.94 23.89
CASP1
25.23 23.59
LOC100859733 23.17 22.14
HSD3B1
27.18 24.76
ALAS1
23.05 19.85
IL16
31.63 27.85
MT4
29.36 26.48
HSP90AB1
24.58 22.86
CD36
22 20.87
SCD
29.16 28.17
LBFABP
19.11
16.3
EEF1A1
16.93 16.44
RPL4
18.92 16.74
TDCPP 50 ug/g
Rep 3 Rep 4
21.13 20.45
24.73 24.49
27.68 28.77
21.95
21.2
22.94 23.45
21.05 20.39
18.5 18.05
22.39 21.92
23.54 23.37
24.69 24.93
14.56 14.33
21.4
20.9
33.33 33.27
33.57 33.89
24.32 24.25
20.58 20.03
24.02 23.84
24.33 24.18
22.7 22.59
25.61 24.45
21.44 21.19
29.06 28.35
28.45
26.2
22.9 23.03
21.22 20.44
28.83 27.86
17.85 16.41
16.35 15.86
17.79 17.33
Rep 5 Rep 6 AVG
SD
21.49 21.11 21.23
0.58
25.56 25.48 25.25
1.40
27.77
27.2 28.04
1.00
22.18
22.1 21.91
0.46
23.93 24.23 23.64
0.71
21.13 22.52 21.50
1.04
18.67 18.62 18.55
0.71
23.81 23.17 23.04
0.86
24.59 24.48 24.15
0.69
25.9 24.57 25.15
1.20
15.43
15.7 14.90
0.87
22.48 22.39 21.79
0.83
36.85 34.93 34.13
0.83
33.28
33.43
0.39
25.27 24.83 24.76
0.63
21.27
22 20.89
1.00
25.12 24.72 24.42
0.57
25.56 25.67 24.76
0.85
23.72 23.59 22.99
0.62
25.56 25.56 25.52
0.95
22.49
23.1 21.85
1.27
29.44 29.37 29.28
1.30
29.53 25.28 27.55
1.80
24.62 24.11 23.68
0.85
21.81 21.65 21.33
0.60
28.77 29.24 28.67
0.55
18.59
18.7 17.83
1.21
17.27 16.61 16.58
0.49
19.21 18.63 18.10
0.97
Supplementary Table 3: Effects of 10 µM bis(1,3-dichloro-2-propyl) phosphate, 24 h exposure
on mRNA expression levels in chicken embryo hepatocytes. Shaded cells indicate a significant
difference in mRNA levels relative to the DMSO control (Two-tailed t-test; p ≤0.05; Fold
change ≥1.7; n=3).
Gene Symbol
CYP3A37
CYP1A4
UGT1A9
SULT1B1
BATF3
PDK4
TXN
ACSL5
HMGCR
SLCO1A2
TTR
DIO1
THRSP
IGF1
NCOA3
SULT1E1
CYP7B1
CASP1
CIDEC
HSD3B1
ALAS1
IL16
MT4
HSP90AB1
CD36
SCD
LBFABP
P-value
Fold Change
0.16
0.02
0.07
0.87
0.11
0.78
0.68
0.23
0.51
0.36
0.08
0.32
0.30
0.31
0.21
0.08
0.11
0.90
0.18
0.06
0.05
0.13
0.01
0.97
0.79
0.19
0.37
1.13
2.34
-1.55
-1.01
-1.17
1.08
1.03
1.14
-1.10
-1.15
1.18
1.10
-1.21
-1.19
-1.17
1.17
-1.47
1.01
1.16
-1.43
1.18
-1.29
-1.62
-1.00
1.04
-1.14
1.12
Supplementary Figure 1: Relationship between fold changes of gene targets on the Avian
ToxChip PCR array measured on embryonic day 18 for the in ovo exposure (50 µg/g TDCPP)
and at 3 (A), 18 (B), 24 (C), and 36 hours (D) for the in vitro (10 µM TDCPP) exposure. The line
of best fit and associated R value are included for each time point.
B.
5
-15
-10
10
R = 0.64
-5
5
10
15
-5
In vitro - 10 M TDCPP
In vitro - 10 M TDCPP
A.
5
-15
-10
-5
5
10
15
-5
-10
-10
In ovo - 50 g/g TDCPP
In ovo - 50 g/g TDCPP
C.
D.
30
R = 0.503
10
5
-15
-10
-5
5
10
-5
In ovo - 50 g/g TDCPP
15
In vitro - 10 M TDCPP
15
In vitro - 10 M TDCPP
R = 0.637
R = 0.512
20
10
-15
-10
-5
5
10
-10
In ovo - 50 g/g TDCPP
15