Supplemental Figure 1
A.
LPS (h):
0
C.
0.5
2
I!B"
ET-1
*
*
6
4
4
2
0
LPS (h)
B.
IκBβ
IκBα
calnexin
LPS (h):
0
D.
0.5
5
mRNA
mRNA
(fold change vs. control)
8
1
IκBβ
IκBα
calnexin
IHMVEC
(fold change vs. control)
THP-1
1
2
I!B"
4
8
ET-1
*
4
3
2
1
0
5
0
5
0
LPS (h)
0
5
0
5
Supplemental Figure 1
LPS induces IκB degradation and expression of NFκB target genes and ET-1 in THP-1 macrophages
(A-B) Representative Western blot showing IκBα and IκBβ in cytosolic extracts from (A) THP-1 and (B) IHMVEC
exposed to LPS (1 µg/mL), with calnexin as loading control. (C-D) Fold change in LPS-induced IκBα and ET-1
gene expression in (C) THP-1 and (D) IHMVEC. Values are means + SEM (n=3/time point); h, hours; *, p <0.05
vs. unexposed control.
Supplemental Figure 2
3
*
*
*
*
*
2
1
WT
AKBI
(fold change vs. control)
Hepatic XOR mRNA
15
*
*
10
*
*
5
*
0
LPS (h) 0 2 4 6
*
0 2 4 6
4
D.
AKBI
*
*
3
2
1
0
LPS (h) 0 2 4 6
0 2 4 6
WT
0 2 4 6
AKBI
2500
Hepatic COX-2 mRNA
0
LPS (h) 0 2 4 6
C.
B.
*
Hepatic FHC mRNA
4
WT
5
(fold change vs. control)
AKBI
(fold change vs. control)
(fold change vs. control)
Hepatic MnSOD mRNA
A.
WT
5
*
2000
1500
*
1000
*
*
* *
500
0
LPS (h) 0 2 4 6
0 2 4 6
Supplemental Figure 2
Inhibition of IκBβ/NFκB signaling does not attenuate LPS-induced NFκB anti- or pro-oxidant target gene expression in
vivo
(A) Hepatic MnSOD mRNA expression in WT and AKBI mice exposed to LPS (50 mg/kg, IP). * = p<0.05 vs. unexposed
controls. Data expressed as means + SE (n=4/time point); h, hours. (B) LPS-induced FHC gene expression in WT and
AKBI mice exposed to LPS (50 mg/kg, IP). * = p<0.05 vs. unexposed controls. Data expressed as means + SE (n=4/time
point); h, hours. (C) LPS-induced XOR gene expression in WT and AKBI mice exposed to LPS (50 mg/kg, IP). * = p<0.05
vs. unexposed controls. Data expressed as means + SE (n=4/time point); h, hours. (D) LPS-induced COX-2 gene
expression in WT and AKBI mice exposed to LPS (50 mg/kg, IP). * = p<0.05 vs. unexposed controls. Data expressed as
means + SE (n=4/time point); h, hours.
Supplemental Figure 3
AKBI
(fold change vs. control)
Hepatic iNOS mRNA
1500
*
*
1000
*
500
0
LPS (h) 0
*
*
*
2 4 6
0 2 4 6
B.
150000
(fold change vs. control)
WT
BMDM ET-1 mRNA
A.
WT
AKBI
*
100000
*
50000
0
LPS (h)
0
1
5
0
1
5
Supplemental Figure 3
Inhibition of IκBβ/NFκB signaling does not attenuate LPS-induced iNOS expression in vivo
(A) Hepatic iNOS mRNA expression in WT and AKBI mice exposed to LPS (50 mg/kg, IP). * =
p<0.05 vs. unexposed controls. Data expressed as means + SE (n=4/time point); h, hours. (B)
LPS-induced iNOS gene expression in WT and AKBI bone marrow derived macrophage following
exposure to LPS (1 µg/ml). Values are means + SEM (n=4/time point); h, hours; *, p <0.05 vs.
unexposed control.