Effects of strong overexpression of DLG using the Gal4 driver C57. A–D, Single confocal slices of third instar larval NMJs stained with antibodies against GTX (green) and DLG (red) in wild type (A, C) and larvae overexpressing DLG (B, D) with the C57 Gal4 driver. E–J, Single confocal slices of an extrasynaptic region of the larval muscles showing the localization of GTX (green) and DLG (red) at the cortical region (E–H) and the subcortical region (I, J) of a muscle cell. K, L, Graphs showing the intensity of the DLG and GTX label at the synapse (K) and the area of GTX-labeled puncta in muscles (L) of wild-type and larvae overexpressing DLG using the C57 Gal4 driver (n = 12 for each genotype). M–O, GTX and DLG immunoreactivity in the regions between the cortical and subcortical DLG networks seen in G–J. *p < 0.05; **p < 0.001. Scale bar: A–D, 5 μm; E–J, 8 μm; M–O, 17 μm.