1. Supplementary Table 1 Akt2 is required for NNK-induced lung tumorigenesis but not for urethane-induced lung tumorigenesis while Akt3 may act as a tumor suppressor for NNK-induced lung tumorigenesis. For each group, littermate mice were injected with 3 weekly doses of 100 mg/kg of NNK and lungs were fixed 16 weeks after the 1st dose. Akt2 littermate mice were injected with 1mg/kg urethane and lungs were fixed after 16 weeks. Lungs from LA2 mice were fixed when the mice reached 8 (Akt1 and Akt2) or 5 (Akt3) weeks of age. All studies were done in a mixed A/J and C57BL/6 background unless otherwise indicated. Surface lung tumors counted Each point represents one mouse.
Supplementary figure 1 Akt1 deletion decreases total Akt in mouse lungs without affecting Akt activation or downstream signaling. Western blots were performed on whole lung extracts from untreated mice. For quantitation, total Akt or pS473-Akt were normalized to ß-actin signal for each sample.

2. Supplementary Table 1 Hollander, et. al
Akt1-/-
Akt2-/-
Akt3-/-
NNK tumor multiplicity
94% decrease in AB6 mixed background*, p=0.0025; 85% decrease in pure A/J, p=0.0001
89% decrease, p<0.0001
2.3X increase, p=0.0107
NNK tumor volume NS
23% decrease in volume, p=0.0161
LA2 tumor multiplicity
57-70% decrease at 5, 8, 12 and 16 weeks, p<0.05 for all
NS at 8 weeks
NS at 5 weeks
LA2 tumor volume
83% decrease at 16 weeks, p<0.0001
NS at 8 weks
4.9-fold increase, p= at 5 weeks
Urethane tumor multiplicity ND
74% increase, p=0.0002
Urethane tumor volume
47% increase, p<0.0001

3. Supplementary Figure 1 Hollander, et. al
Akt signaling in normal lungs
+/ Akt1+/ Akt1-/ Akt2-/- Akt3-/-
Akt
pS473-Akt
Akt1
Akt2
Akt3
pS6
ß-actin
Figure S2 Akt1 deletion decreases total Akt in mouse lungs without affecting Akt activation or downstream signaling. A, western blots on whole lung extracts from untreated mice., B, quantitation of A and p values. 1, 2 and 3 are Akt1, Akt2 and Akt3 genotype.