Fig. 2: In vivo activity of cyclic fasting and CBIs against PDAC and CRC xenografts.

a–e Capan-1 xenografts were established in 6-8-week-old female athymic nude mice. Once tumours were palpable, mice were randomized to be treated with ad libitum diet (n = 12), weekly 48 h water-only fasting (“fasting”; n = 12), terbinafine (40 mg/kg/day for five days a week; n = 10) or combined fasting plus terbinafine (n = 12). Tumour volume was monitored at the indicated time points (a). At the end of the experiment, the excised tumours were weighted (b) and used for cholesterol content determination (c). Peripheral blood was collected before treatment onset and at the end of the experiment and serum was used to detect total cholesterol (d), HDL and LDL cholesterol (e) by standard biochemistry. HDL and LDL cholesterol concentration was used to determine the HDL/LDL ratio (e). HCT116 xenografts were established in 6-8-week-old female athymic nude mice. Once tumours were palpable, mice were randomized to be treated with ad libitum diet (n = 7 in (f); n = 8 in h), weekly 48 h water-only fasting (n = 9 in f; n = 6 in h), simvastatin (in f, 80 mg/kg/day; n = 7), combined fasting plus simvastatin (in f, n = 10) clotrimazole (in h, 60 mg/kg twice a week; n = 7), combined fasting plus clotrimazole (in h, n = 8), or fasting, clotrimazole and human LDL (in h, 0.25 mg/mouse twice a week coupled with fasting; n = 6). Tumour volume was monitored at the indicated time points (f, h). At the end of the experiment, tumours were weighted (g, i), imaged (g) and utilized for intratumour cholesterol determination (j). In (a, f, h), n indicates the number of tumours per treatment group. In (b, c–e, g, i, j), data points are biological replicates: they represent single tumours (b, c, g, i, j) or sera from different animals (d, e). Data are shown as mean ± SEM in (a, b, f, g, h, i) and as mean ± SD in (c–e, j). In (a, c, h, j), data were analysed by one-way ANOVA with Tukey post-test. In (b, d, e, f, g, i), p values were calculated by two-tailed Student’s t test. In (f), the statistical analysis was performed utilizing the tumour volumes from day 24. Source data are provided as a Source Data file.