DNRX interacts with Scribble and sustains the terminal SV aggregation.
A–B‴, representative images of larvae brains of the indicated genotypes were stained with DNRX (red), HRP (green), and DAPI (blue), showing the level of DNRX was decreased in scribble mutant. C–D″, representative images of larvae NMJ of the indicated genotypes were stained with DNRX (green) and HRP (red), showing the level of DNRX was decreased in the scribble mutant. E, quantification of the fluorescence intensity for DNRX both in brain and NMJ of the indicated genotypes. F and G, Western blot analysis of protein lysates prepared from heads using anti-DNRX antibody showing that the level of DNRX was reduced in scribble mutant both in larvae (F) and adult (G) stages. H, quantitative analysis of data for Western blot analysis of the indicated genotypes for adult flies. I, Western blot analysis of protein lysates prepared from heads using anti-GFP antibody showing that the level of Scribble was reduced in the dnrx mutant in larvae stage. J, quantitative analysis for Western blots of the indicated genotypes. K–L″, representative images of larvae brain of the indicated genotypes stained with DNRX (red) and GFP (green), showing the level of Scribble was decreased in dnrx mutant. M–N‴, representative images of larvae NMJ of the indicated genotypes were stained with HRP (blue), DLG (red), and GFP (green), showing the level of Scribble was decreased in dnrx mutant. O, quantification of the fluorescence intensity for Scribble::GFP of the indicated genotypes. P–R″, synaptic boutons of wild-type (P–P″), dnrx mutant (Q–Q″), and pre-synaptic overexpress Scribble in dnrx mutant (R–R″) double stained for SYT (red) and HRP (green), which labels the pre-synaptic SVs and neuronal membrane, respectively. S, quantification of the ratio of SYT-diffused boutons in muscle 4 shows that dnrx mutants have disturbed the terminal cluster of SVs and this phenotype can be rescued by pre-synaptic Scribble. Data are mean ± S.E. ***, p < 0.001; **, p < 0.01; and *, p < 0.05. Two-tailed Student's t tests were used to compare genotypes. Scale bar, 50 (A–B‴), 5 (C–D″), 50 (K–L″), 20 (M–N‴), and 5 μm (P–R″).