Fig 10. Pharmacodynamics in 4T1 tumours.

(A, B) Female Balb/c mice (n = 4) bearing 4T1 tumours were treated with 40 mg/kg of AL776, 20 mg/kg each of gefitinib + dasatinib or vehicle and sacrificed 1h and 24h after drug administration (intravenous, i.v). Tumours were collected and western blot analysis was used to detect inhibition of phosphorylation of EGFR, c-Src and ERK1/2 by the drug, at different time points. The bands were quantified and represented as histograms and as a ratio of phospho-protein/total protein. Statistical significance was determined using multiple t-test and p < 0.05 was considered significant. (C) Efficacy study was carried out in female Balb/c mice (n = 6) bearing 4T1 tumours and treated with 40 mg/kg of AL776 or vehicle administered IV (once/day) for 5 consecutive days. Tumour growth was monitored for two weeks by measuring tumour volume on alternate days. Graphs representing the average tumour volumes ± SEM of the two groups are shown. (D) On the last day of the study, tumours were collected and their average ± SEM weight (g) from AL776 treated and the control groups (n = 6) are plotted as histograms.