Figure 5. mnb mutant phenotype can be rescued by TOR activation in flies.

(A–F and J–L) Third instar larval neuromuscular junction (NMJ) (muscles 6/7) were stained using anti-HRP (Green) and anti-Dlg (Red). Muscles are stained with phalloidin (Blue, A–F). HRP (green) stains the entire neuron and Dlg (red) stains only boutons (Red + Green). (G–I, M) Quantification of bouton numbers, normalized to muscle area (Bouton-NMA). Error bars represent standard deviation. Statistical significance (p-values: ***<0.001; **<0.01; *<0.05) is calculated by unpaired student’s t-test. (A, B, G) mnb¹ alleles show fewer boutons numbers as compared to wild-type (WT, Canton S) control (B). Data are quantified in G. (C, D, H) mnb overexpression (D42-Gal4>UAS mnb, D) increases bouton numbers as compared with mCherry overexpression (D42-Gal4>UAS-mCherry, Control, C). D42-Gal4 is a motor-neuron-specific driver. Data are quantified in H. (E, F, I) Rheb overexpression (D42-Gal4>UAS Rheb, F) increases bouton numbers as compared with mCherry overexpression (D42-Gal4>UAS-mCherry, Control, E). Data are quantified in I. (J–M) Rheb overexpression in mnb mutant (mnb¹/Y D42-Gal4>UASRheb, L) suppressed bouton phenotype as compared to mnb mutant (mnb¹/Y D42-Gal4/+, K). Wild-type is heterozygous D42-Gal4 (+/Y; D42-Gal4/+, J). Data is quantified in (M).

Figure 5—figure supplement 1. Neuromuscular junction (NMJ) phenotypes due to TOR gain or loss.

NMJ (muscles 6/7) are stained using anti-HRP (Green) and anti-Dlg (Red). Muscles are stained with phalloidin (Blue, A–F). HRP (green) stains the entire neuron and Dlg (red) stains only boutons (Red + Green). (G–I, M) Quantification of bouton numbers - normalized to muscle area (Bouton-NMA). Error bars represent standard deviation. Statistical significance (p-values: ***<0.001; **<0.01; *<0.05) is calculated by unpaired student’s t-test. (A–C) gig¹⁰⁹ alleles show increased bouton numbers (B) as compared to wild-type (WT, Canton S) control (B). Data is quantified in C. (D–F) Expression of dominant negative TOR (D42-Gal4>UAS TOR.ted, E) decreases bouton numbers as compared to control (D42-Gal4/+, D). D42-Gal4 is a motor neuron-specific driver. Data is quantified in F.