FIGURE 7.

Cigarette smoke reduces Akt and p-Akt and induces ttc3 expression and Akt ubiquitination in rat lungs. Lewis male rats were exposed to cigarette smoke (SM) or clean room air (CON) for 3 months. A, induction of ttc3 expression. Expression of ttc3 and gapdh was assessed by RT-PCR. B, reduction in Akt and p-Akt and induction of Akt ubiquitination in rat lungs. Akt ubiquitination in rat lungs was confirmed by two methods, ubiquitinated protein pulldown (Ubi pull-down) and immunoprecipitation of Akt (IP: Akt), followed by Western blot (WB) with anti-Akt and anti-Ubi antibodies, respectively. Akt and p-Akt levels were determined before pulldown or immunoprecipitation (IP) (Input). Asterisks denote a statistically significant difference (p < 0.05) between the control and smoke groups. Number of each group is five.