Figure 3

1NA-PP1-induced SAD-1 as inactivation is dose-dependent. (A) Dose-response in polarity phenotype. L123A SAD-1-expressing sad-1 animals were exposed to various concentrations of 1NA-PP1, and the number of ectopic juIs1 puncta on the dorsal side of each animal was counted. At 4 μM, the level of polarity defect was similar to the animals exposed to DMSO. At 8 μM and 17 μM, the inhibiting effects were stronger but incomplete. At 33 μM, complete inhibition of SAD-1as was observed. 1NA-PP1 had no effect on the polarity phenotype of wild-type (wt) animals. N.S., not significant, Wilcoxon rank-sum test. Error bars indicate standard deviations. (B, C) Dose-response in synaptic organization phenotype. At 33 μM, 1NA-PP1 had no effect on the juIs1 morphology of wild-type animals (compare Bi and ii; quantified in Biii). L123A SAD-1-expressing sad-1 animals were exposed to various concentrations of 1NA-PP1, and the juIs1 morphology on the dorsal side of the animals was observed (Ci). At 4 μM and 8 μM, most animals appeared normal. At 17 μM, while some juIs1 puncta appeared defective (upper panel), others were normal (lower panel). At 33 μM, the rescuing effects of SAD-1as were inhibited in all animals. Quantification and density estimates of the punctum widths show that while 4 μM 1NA-PP1 had little effect (Cii), 33 μM 1NA-PP1 completely inhibited L123A SAD-1 (Ciii). All density graphs show punctum widths on the x-axis and density on the y-axis. Scale bar, 5 μm.