Fc1
Apcfl/+
Pik3cap110* CRC spheroids possess a lox-stop-lox sequence prior to the p110* transgene, which results in a constitutively active PI3K (A and B). These spheroids were allowed to mature for 24–96 hours prior to treatment with BEZ235, TAK-228 or control (C and D). Baseline and 48 hours post-treatment brightfield images were obtained and change in spheroid diameter measured for individual spheres. A similar median sphere size following treatment with BEZ235 and TAK-228 were observed and both significantly reduced from control (p < 0.001) (E). Reduced phosphorylation of AKT (ser473), RPS6 (Ser235/236), and 4EBP1 (Thr37/46) was observed after treatment of these spheres with BEZ235 and TAK-228 compared to control (F). Population distribution histograms demonstrating variation in change in sphere size for Fc1
Apcfl/+
Pik3cap110* CRC spheroids after 48 hours of treatment with BEZ235, TAK-228 or control (G). Size bar in panel A = 1 mm.