Synaptic vesicles are mislocalized at jaguar mutant boutons as revealed by immunostaining against synaptotagmin. A representative control Ib bouton and mutant jar322/Df(3R)crb87-5 Ib bouton are shown with their corresponding plot profiles (A and B). There was a significant reduction in relative difference in fluorescence intensity across jar322/Df(3R)crb87-5 mutant boutons (n = 13) compared to the control boutons (n = 25; Unpaired T-test, *** = p < 0.001) (C). Mean fluorescence intensity did not differ significantly between the mutant (n = 48) and the control (n = 64; Unpaired T-test, p > 0.05) (D). Diffuse jar322/Df(3R)crb87-5 boutons (n = 53) were significantly larger than control boutons (n = 64) and jar322/Df(3R)crb87-5 boutons with a normal, doughnut-shaped distribution of synaptotagmin (n = 27, Dunn's Multiple Comparison Test, *** = p < 0.001) (E). Boutons used for this study were from muscle 6/7 NMJs in segments A3, A4 and A5. Bars represent mean ± SEM.