Figure 9.

Pharmacological Inhibition of PI3K, ERK, and Wnt/β-Cat Signaling Partially Rescues Polyploidy in AhR−/− Liver

(A and B) Primary hepatocytes from AhR+/+ (A) and AhR−/− (B) mice were cultured for 48 hr in the presence of inhibitors for PI3K (LY294002), ERK (PD98059), and Wnt/β-Cat (salinomycin) at a low or high concentration (see Methods). Their DNA content was analyzed by flow cytometry and represented as 2c, 4c, and >4c containing cells.

(C) AhR−/− mice were treated in vivo with those inhibitors (see Methods), and the activation of PI3K signaling determined by the levels of p-AKT^Ser473 with respect to total AKT expression.

(D) mTOR expression was also determined in vivo under the same experimental conditions indicated above.

Two representative mice for each condition are shown. Six mice of each genotype were perfused to isolate primary hepatocytes for in vitro experiments and five mice were treated with inhibitors in vivo. Three technical replicates were analyzed. Data are shown as mean ± SD. Scale bar, 15 μm. Antibodies and oligonucleotides used are indicated in Tables S1 and S2, respectively. SD, standard deviation.