All Drosophila stocks and crosses were maintained at 25°C.
Immunohistochemistry was done using a protocol described previously (Varelas et al.,
2008 ). The primary antibodies were rabbit anti-Yki (1:400; a gift from K. Irvine), mouse anti-DIAP1 (1:100; a gift
from B. Hay), rabbit anti-b-gal (1:100; Invitrogen), and rabbit anti-dMyc (1:100; Santa Cruz Biotechnology). The secondary
antibodies were donkey anti-mouse or anti-rabbit IgG conjugated to Cy3 (1:200; Jackson ImmunoResearch). Samples were imaged
using an Olympus Fluoview 1000 confocal microscope, and the images were edited using Adobe Photoshop CS6.0. Brain lobe size in
pixels was measured using the histogram function of Adobe Photoshop CS6.0. Data are presented as the means and standard
deviations calculated for each sample. A 2-tailed t test performed with Excel 2013 was used to determine whether the observed
differences were significant (p < 0.05).
Immunohistochemistry was done using a protocol described previously (
2008
from B. Hay), rabbit anti-b-gal (1:100; Invitrogen), and rabbit anti-dMyc (1:100; Santa Cruz Biotechnology). The secondary
antibodies were donkey anti-mouse or anti-rabbit IgG conjugated to Cy3 (1:200; Jackson ImmunoResearch). Samples were imaged
using an Olympus Fluoview 1000 confocal microscope, and the images were edited using Adobe Photoshop CS6.0. Brain lobe size in
pixels was measured using the histogram function of Adobe Photoshop CS6.0. Data are presented as the means and standard
deviations calculated for each sample. A 2-tailed t test performed with Excel 2013 was used to determine whether the observed
differences were significant (p < 0.05).