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.

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=0.0177 at 5 weeks Urethane tumor multiplicity ND 74% increase, p=0.0002 Urethane tumor volume 47% increase, p<0.0001

Supplementary Figure 1 Hollander, et. al Akt signaling in normal lungs +/+ Akt1+/- Akt1-/- Akt2-/- Akt3-/- 1 2 3 4 1 2 3 4 1 2 3 4 1 2 1 2 3 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.