Fig. 6 | Nature Communications

Fig. 6

From: Survival of pancreatic cancer cells lacking KRAS function

Fig. 6

PI3K inhibition enhances cap-dependent translation inhibition in KRAS deficient cells. a Schematic of cap-dependent translation reporter construct. 5′-LTR 5′ long terminal repeat of MSCV virus with promoter activity. In transduced cells, mCherry expression correlates with cap-dependent translation and GFP expression correlates with cap-independent translation initiated via an internal ribosomal entry site (IRES). b FACS plots of GFP and mCherry fluorescence in KRAS intact and deficient (KO) cells. KRAS deficient cells exhibited a greater decrease in mCherry (relative to GFP) expression when treated for 24 h with GDC-0941 (2 μM) or the mTORC1/2 inhibitor AZD8055 (100 nM) than intact cells. Triangle gates were drawn along the midline diagonal of the FACS plots of DMSO-treated cells and maintained in plots of drug treatment. Numbers denote percentages of cells within gate and is inversely related to cap-dependent translation of reporter. c FACS plots of GFP and mCherry fluorescence in KRAS deficient cells transduced with myr-AKT1 or myr-AKT1 (K179M). Wild-type AKT1 expression decreased the effect of GDC-0941 on cap-dependent translation compared to its kinase-dead variant