Figure 3: Gcm^R59L overexpression increases glial cell number less efficiently than Gcm.

(a–d”) Confocal projections of Stage 15 embryos immunostained with HRP (magenta) to reveal neuronal membranes and Repo (green) for glia for the following genotypes: (a–a”) Sca-Gal4 > w¹¹¹⁸ (Control), (b–b”) Sca-Gal4 > UAS-gcm, (c–c”) Sca-Gal4 > UAS-gcm^R59L-2, and (d–d”) Sca-Gal4 > UAS-gcm^R59L-3. Scale bar: 100 μm. (e) Numbers of Repo-positive cells per hemisegment were scored from the midline (yellow arrow) to the 5 LCH-associated glia (yellow arrowheads). (f) Numbers of glia per hemisegment in Stage 15 were shown for embryos with the above genotypes. Repo-positive cells were count for hemisegments A1–A4, with n = 24 (w¹¹¹⁸), n = 48 (UAS-gcm), n = 30 (UAS-gcm^R59L-2), and n = 40 (UAS-gcm^R59L-3). (g) Luciferase reporter assays were performed for cells transfected with p3xFlag-gcm or p3xFlag-gcm^R59L in the absence or presence of MG132 (50 μM). Note that a decrease in the activity was seen for Gcm^R59L. In the presence of MG132, however, no difference was detected for either form. Averages are mean ± SEM, * represents p < 0.05, ** represents p < 0.01, and *** represents p < 0.001 by Student’s t test between two groups and one-way ANOVA test among multiple groups. ns means no significance.