Research Paper Volume 13, Issue 21 pp 24017—24036

Low-protein diet applied as part of combination therapy or stand-alone normalizes lifespan and tumor proliferation in a model of intestinal cancer

Afatinib reduces the EgfrCA-induced phenotype. Control animals (esg > w1118) and animals with Egfr-induced over-proliferation (esg > EgfrCA) of intestinal stem cells and enteroblasts (esg+ cells) were treated with afatinib (100 μM) during induction. The esg+ cells are marked with GFP. (A, B) Intestines of control flies with and without treatment after 5 days and (A’, B’) 15 days. (C, D) Intestines of EgfrCA animals with and without treatment after 5 days and (C’, D’) 15 days. (E) Quantification of the area covered by GFP-positive cells in midguts indicating the number of esg+ cells after 5 days of intervention. n = 10–13. (F) The lifespan of animals treated with 100 μM or 50 μM afatinib. n = 40. Statistical significance was tested by one-way ANOVA and the Tukey test. Lifespan significance was tested by the log-rank (Mantel-Cox) test. Significances are marked with lines or corresponding color. ns = not significant, ** = p *** = p **** = p

Figure 2. Afatinib reduces the EgfrCA-induced phenotype. Control animals (esg > w1118) and animals with Egfr-induced over-proliferation (esg > EgfrCA) of intestinal stem cells and enteroblasts (esg+ cells) were treated with afatinib (100 μM) during induction. The esg+ cells are marked with GFP. (A, B) Intestines of control flies with and without treatment after 5 days and (A’, B’) 15 days. (C, D) Intestines of EgfrCA animals with and without treatment after 5 days and (C’, D’) 15 days. (E) Quantification of the area covered by GFP-positive cells in midguts indicating the number of esg+ cells after 5 days of intervention. n = 10–13. (F) The lifespan of animals treated with 100 μM or 50 μM afatinib. n = 40. Statistical significance was tested by one-way ANOVA and the Tukey test. Lifespan significance was tested by the log-rank (Mantel-Cox) test. Significances are marked with lines or corresponding color. ns = not significant, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001. Scale bar: 100 μm.