Synaptotagmin expression is unaltered in phospholipid-binding mutants. A, Synaptotagmin is expressed at similar levels in each of the transgenic synaptotagmin lines. Representative Western blots of homogenized CNSs of third instars from the indicated lines were probed with an anti-synaptotagmin antibody. To confirm equal loading, they were also probed with an anti-actin antibody. B, Comparison of synaptotagmin/actin ratio normalized to the mean ratio of the transgenic control for P[sytWT](n = 45), P[sytA-RQ] line 7 (n = 6), P[sytA-RQ] line 8 (n = 14), P[sytB-RQ] line 3 (n = 18), and P[sytB-RQ] line 4 (n = 10). An ANOVA showed no significant difference between any of the genotypes (p > 0.1). C, Synaptotagmin is localized to the larval neuromuscular junction in each of the transgenic synaptotagmin lines. Representative confocal Z-stack projections are shown for P[sytWT], P[sytA-RQ] line 8, and P[sytB-RQ] line 4. Scale bar, 10 μm.