Depletion of neuronal NMII ^HC reduces the levels of the NMII ^LC subunit in the muscle. (A) Distribution of Sqh::GFP at NMJs in fixed larvae expressing CTRL³⁵⁷⁸⁵, Zip⁶⁵⁹⁴⁷, and Zip³⁶⁷²⁷ RNAi in neurons (C155-Gal4). Sqh::GFP fluorescence was analyzed within the α-HRP–delineated NMJ area (inner yellow outline), as well as 1 µm outside the neuron in the postsynaptic area (between inner and outer yellow outline). (B) Analysis of the Sqh::GFP fluorescence in the HRP-delineated, postsynaptic, and muscle ROIs. (C) Example of WEKA-segmented individual Sqh::GFP particles in the HRP-delineated and postsynaptic compartments. (D) Analysis of the number, area, and fluorescence WEKA-segmented sqh::GFP particles within the HRP-delineated NMJ area. (E) Analysis of the number, area, and fluorescence of postsynaptic WEKA-segmented sqh::GFP particles. Box-and-whisker graphs were used to represent the results of the area and fluorescence intensity of the individual Sqh 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. N in bar graphs—NMJs and muscle ROIs; N in box and whiskers—individual Sqh particles. **P < 0.01, ***P < 0.001 after one-way ANOVA with Šídák’s multiple comparisons test. g—Hedges’ g represents the effect size. Scale bar—2 µm. See Table S1 for detailed genotypes and N.