Extended Data Fig. 3. Wnt-signalling is still active after Raptor deletion and Rapamycin treatment causes regression of established tumours.

a+b) Representative IHC of MYC and β-catenin showing high MYC levels and nuclear localisation of β-catenin 96hrs after Apc and Apc/Raptor deletion, demonstrating active Wnt-signalling. Nuclear staining (as opposed to membranous staining) of β-catenin is indicative of active Wnt-signalling; Scale bar = 100μm (representative of 3 biological replicates). c) Kaplan-Meyer survival curve of Apc^Min/+ mice treated with rapamycin when showing signs of intestinal neoplasia. 10mg/kg rapamycin treatment started when mice showed signs of intestinal disease, and was withdrawn after 30 days. Animals continued to be observed until signs of intestinal neoplasia. Death of animals in the rapamycin group almost always occurred following rapamycin withdrawal. (n=8 per group, ***p-value≤0.001, Log Rank test); d) boxplot showing that 72hr 10mg/kg rapamycin treatment causes an increase in Lysozyme positive cells in tumours. Percent Lysozyme positivity within tumours was calculated using Image J software (http://imagej.nih.gov/ij/). Whiskers are max and min, black line is median (10 tumours from each of 5 mice per group were measured, **p-value≤0.014, Mann-Whitney U test); e) boxplot showing that 72hrs 10mg/kg rapamycin treatment causes a decrease in BrdU positivity within tumours. Percent BrdU positivity within tumours was calculated using Image J software. Whiskers are max and min, black line is median (10 tumours from each of 5 mice per group were measured, **p-value≤0.021, Mann-Whitney U test); f) Representative IHC of Lysozyme, showing a lack of Lysozyme positive paneth cells in remaining cystic tumours after 30 days of 10mg/kg rapamycin treatment; Scale bar = 100μm (representative of 5 biological replicates).