Fig. 1

Reduced expression of Not3 increases tumor formation in Drosophila melanogaster eye cancer models. a Qualitative and quantitative representation of the tumor burden upon downregulation of Not3 on sensitized flies. Bars show the percentage of eyes screened: blue, normal eyes; orange, hyperplastic eyes; *** p < 0.001. Three independent experimental crosses were established. The mean number of eyes screened is shown on the graph (Y-axis). Microscopy images show eyes of adult flies from representative genotypes, scale bars are 200 μM. b qPCR analyses showing the Not3 expression levels from each genotype. Expression values are calculated from three independent crosses. From each cross, 30 eye-antennal imaginal discs were dissected. The red dotted bar represents the expression value = 1. c Quantification of BrdU and pH3 positive cells on the posterior portion of eye-antennal imaginal discs. Values for 10 individual discs were plotted. Representative confocal images of eye-antennal imaginal discs of the indicated genotypes. Green staining: BrdU or pH3 positive cells; red staining: ELAV protein. Scale bars are 70 μM; ns, not significant; * p < 0.05; *** p < 0.001. d Qualitative and quantitative representation of the tumor burden upon downregulation of Not3 on the Ras-V12 background. Bars show the average percentage of eyes screened: light orange, eyes classified as score 1 (presence of 1 fold); yellow, eyes classified as score 2 (presence of 2–4 folds); dark orange, eyes classified as score 3 (presence of more than 4 folds); *** p < 0.001. Three independent experimental crosses were established. The mean number of eyes screened is shown on the graph (Y-axis). Microscopy images show eyes of adult flies from representative genotypes, scale bars are 200 μM. e Quantitative representation of the whole eye area (region with presence of differentiated photoreceptors) on representative adult eyes (n = 8) with downregulation of Not3 on the Ras-V12 genetic background. Control eyes measured values were considered as 100%; ** p < 0.01