Fig. 6: Fasting and CBIs inhibit AKT and STAT3 signalling in cancer cells.

a Tumours (Capan-1 xenografts) were isolated at the end of the experiment presented in Fig. 2a and utilized for protein lysate generation and for the detection of phosphorylated (Ser 473) and total AKT, phosphorylated (Tyr705) and total STAT3 by Western blotting. The intensity of the phosphorylated AKT and STAT3 bands were quantified and normalized to that of the total AKT and STAT3 bands, respectively. b Capan-1 cells were treated for 24 h w/ or w/o FMCC, combined IGF1 (5 ng/ml)+leptin (50 ng/ml)+insulin (500 pM) (“fasting-reduced factors” - FRFs- combination). Thereafter, 3.5 mM methyl-β-cyclodextrin (MβCD) w/ or w/o FMCC was added for 3 h where indicated. Afterwards, MβCD was replaced by 20 μM terbinafine for 24 h where indicated. Finally, cells were used for protein lysate generation and phosphorylated (Ser473) and total AKT, as well as phosphorylated (Tyr705) and total STAT3 were detected by immunoblotting. c Capan-1 cells were transduced with a constitutively active form of STAT3 (STAT3-C), with myr-AKT or with both STAT3-C and myr-AKT. Thereafter, cells were treated for 96 h w/ or w/o FMCC and during the last 72 h w/ or w/o 25 μM terbinafine. In the Western blot band quantifications presented in (a), samples derive from the same experiment; blots were processed in parallel; data points are biological replicates (they represent single tumours). In (b), one out of two independent experiments is presented. In (c), data points are experimental replicates. In (a, c), data are shown as mean ± SD. p values were calculated by two-tailed Student’s t test. Source data are provided as a Source Data file.