Depletion of neuronal NMII induces rearrangements of NMII ^HC /Zip pre- and postsynaptically. (A) Endogenous NMII^HC/Zip (cyan) at NMJs (HRP—magenta) in fixed larvae expressing CTRL³⁵⁷⁸⁵, Zip⁶⁵⁹⁴⁷, and Sqh³²⁴³⁹ RNAi in neurons (C155-Gal4); white arrows—Zip puncta; gray arrows—Zip aggregate-like puncta. (B) Quantification of Zip fluorescence measured between the presynaptic area (masked by the neuronal HRP signal, inner yellow outline) and 1 µm outside the neuron in the postsynaptic area (outer yellow outline), along with analysis of the number, area, and fluorescence of WEKA-segmented individual Zip particles in this postsynaptic compartment. N in bar graphs—NMJs; N in box and whiskers—individual Zip assemblies. (C) Quantification of Zip fluorescence in different muscle ROIs, along with analysis of the number, area, and fluorescence of WEKA-segmented individual Zip aggregate-like structures in muscles. N in bar graphs—NMJs or muscle ROIs; N in box and whiskers—individual Zip particles. Box-and-whisker graphs were used to represent the results of the area and fluorescence intensity of the individual Zip particles. Whiskers represent 10^th to 90^th percentile, while the rest of the data point are shown as individual values. The y axis in these graphs represents log₁₀, to capture the broad distribution of the individual values. In bar graphs with linear y axis, error bars represent the SEM. **P < 0.01, ***P < 0.001 after one-way ANOVA with the Kruskal–Wallis multiple comparisons test. g—Hedges’ g represents the effect size. Scale bar—2 µm. See Table S1 for detailed genotypes and N.