Hollander, et al, Table S1

1
Hollander, et al, Table S1
Table S1 Decreased NNK-induced O6-mG adduct
formation in the lung but not liver of pseudo-AJ
Akt2-/- mice. Akt2/ and Akt2-/- mice were
injected with NNK at a dose of 200 mg/kg, and
levels of O6-mG, as well as levels of total
guanine, in the liver and lung were determined at
4 h after the injection. Data represent means
S.D. (n 6 3 each male and female).
2
Hollander, et al, Table S2
Table S2 In vitro metabolism of NNK by lung and
liver microsomes from Akt2/, Akt2/- and
Akt2-/- mice in a pure A/J background. Rate of
formation of three major NNK metabolites (HPB,
NNK-N-oxide, and sodium bisulfite-trapped OPB)
were determined. Reaction mixtures contained 100
mM potassium phosphate buffer, pH 7.4, 10 µM NNK,
0.20 mg/ml lung or liver microsomal protein from
three strains of mice (3 each male and female), 5
mM sodium bisulfite, and 1.0 mM NADPH. The
values presented are means S.D. (n 6).
3
Hollander, et al, Figure S1
Figure S1 Equal CYP2A protein in lung and liver
microsomes from pure A/J mice Akt2/ and Akt2-/-
mice . Lung (5 µg protein) and liver (1 µg
protein) microsomes pooled from 3 mice of each
strain were analyzed (same samples as used for
table S2). Densitometry analysis indicated that
maximal difference in band intensity among the
three strains is less than 5 of the averaged
values for either liver or lung.
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Hollander, et al, Table S3
Table S3 Decreased NNK-induced O6-mG adduct
formation in the lung but not liver of Akt2-/-
and Akt2/- mice in a pure A/J mice bakcground.
Akt2/, Akt2/- and Akt2-/- mice were injected
with NNK at a dose of 100 mg/kg, and levels of
O6-mG, as well as levels of total guanine, in the
liver and lung were determined at 4 h after the
injection. Data represent means S.D. (n 6 3
each male and female).
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Hollander, et al, Table S4
Table S4 In vitro metabolism of NNK by lung and
liver microsomes of A/J and 129/Sv mice Rate of
formation of three major NNK metabolites (HPB,
NNK-N-oxide, and sodium bisulfite-trapped OPB)
were determined. Reaction mixtures contained 100
mM potassium phosphate buffer, pH 7.4, 10 µM NNK,
0.20 mg/ml lung and liver microsomal protein from
two strains of mice, 5 mM sodium bisulfite, and
1.0 mM NADPH. The values presented are means
S.D. (n 3).
6
Hollander, et al, Figure S2
Figure S2 Decreased urethane-induced lung tumor
multiplicity in Akt2-/- in a A/J-N8 background. 6
week old mice were injected with 1 mg/kg
urethane, intraperitoneally. Lungs were harvested
16 weeks after treatment. Each point represents
one mouse.
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Hollander, et al, Figure S3
A/J CYP2A5 129 CYP2A5 CYP2A6 CYP2A13
Figure S3 Alignment of mouse CYP2A5 (A/J and 129
strains) and human CYP2A6 and CYP2A13. Shading
indicates amino acid identity. Numbers at the
top denote amino acid position within the
protein. Red arrow indicates amino acid 117.
The A/J CYP2A5 (117A) is similar to human CYP2A13
(117A), while the 129 CYP2A5 (117V) is similar to
the human CYP2A6 (117V).