(A) Time course analysis of effects of LY294002 (LY) on SP10 mouse lung tumour cell line. Shown are Western blots of cell lysates probed with antibodies to p-Akt, p-FOXO, and p27 at the indicated times after addition of serum and LY. The control (ctrl) lane is from cells not serum starved. (B) Real-time PCR of p27 mRNA from treated (+LY) and control (−LY) treated SP10 cells 8 and 24 h after start of PI3K inhibition. (C) Western blot of total and p-Akt, p-FOXO, and p27 in lung tumours from wild type mice 5 h after treatment with LY. n: nuclear extracts, c: cytoplasmic fractions. (D) Sections of representative lung tumours immunostained for p27 shows increase in nuclear p27 following LY294002 treatment. (E) Western blot of tumour lysates shows decreased p-T198 p27 and p-S10 p27 in LY294002 treated lung tumours. Lamin B1 is nuclear fraction loading control. N.Lung = normal lung.