Figure 5.

Chloroquine and rapamycin exert additive effects on the inhibition of large MCF-7 tumour growth. (A) Representative immunofluorescence CAIX (green) and LC3 (red) staining of MCF-7 tumour sections harvested from the vehicle group at different tumour diameters (5–7, 7–10 and >10 mm); 4′,6-diamidino-2-phenylindole (DAPI) counterstaining (blue) is also shown. Insets show magnification of LC3 punctate staining. (B) Graphs represent the effects of intraperitoneal administration of rapamycin (1 mg kg⁻¹ per day), CQ (50 mg kg⁻¹ per day) or both on the growth of MCF-7 tumours injected in the mammary fat pad. Treatment was initiated when tumours reached a diameter of 10 mm; mice treated with vehicle were used as controls (*P<0.05, ***P<0.001, n=10 mice per group). (C) Bar graphs show the quantification of CAIX immunofluorescent signals in the different conditions (*P<0.05, **P<0.01, ***P<0.001, n=5). (D) Typical haematoxylin colouration of MCF-7 tumour sections harvested from the rapamycin and the rapamycin-CQ groups as described above. Bar graphs represent the extent of the central necrosis area (*P<0.05, n=4).