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Fig. 6 | Hereditas

Fig. 6

From: Overexpression of the Drosophila ATR homologous checkpoint kinase Mei-41 induces a G2/M checkpoint in Drosophila imaginal tissue

Fig. 6

Response of the p53R-GFP reporter on mei-41 overexpression. (a) In response to p53 activation, nuclear GFP is expressed from the p53R-GFP reporter [36]. This system was used to assay p53 activation in consequence of the overexpression of either lok, mei-41 or grp; lacZ served as control. Salivary glands were analyzed; their nuclei visualized with the nuclear marker Pzg [39]. In contrast to lacZ and grp, overexpression of lok and to a lesser degree mei-41 resulted in a robust induction of the p53R-GFP reporter. Size bar represents 100 μm in all panels. (b) Quantification of nuclear p53R-GFP intensity was determined relative to the mean intensity of the nuclear marker protein Pzg (n = 84). Induction of lok strongly induced p53R-GFP nuclear accumulation. Also mei-41 caused a significant nuclear accumulation of p53R-GFP, whereas grp did not. *** p < 0.001; ** p < 0.01; ns. not significant according to ANOVA two tailed Dunnet’s approach. (c) Expression levels of GFP were quantified by qRT-PCR in third instar larvae. Overexpression of lok and mei-41 with da-Gal4 considerably increased p53R-GFP reporter gene activity in relation to the lacZ-control: lok 27.5-fold, mei-41 4.9-fold and grp 1.6-fold. Data were assembled from four biological and two technical replicates. Mini-max depicts 95% confidence, median corresponds to expression ratio. As reference genes, cyp33 and Tbp were used. Efficiencies for GFP (0.91), for cyp33 (0.96) and Tbp (0.95) were accounted for in determining relative quantities [44]

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