Fig. 7: Functions and properties of the neuromuscular junction (NMJ) model derived from the coculture of SOD1^D90D and SOD1^D90A motor neurons (MNs) and SOD1^D90D skeletal muscles (SKM).

A 3D confocal images of immunocytochemistry (ICC) staining for the SKM marker TTN; neuronal marker NF; and NMJ markers SV2 and AChR. B–F Calculations of the volume of SKM (B), MN (C), and the ratios of MN/SKM (D), AChR/SKM (E), and AChR/MN (F) in all cross-coculture NMJ models. For each group, 5 images were analyzed from 3 independent experimental replicates, with each data point representing the mean value of a single replicate. G Bright-field intensity changes in different ROIs after CaCl₂ induction in coculture NMJs. H Spontaneous and CaCl₂-induced SKM contraction ratios in NMJs. In total, 19 (SOD1^D90D MN + SOD1^D90D SKM), 30 (SOD1^D90A MN + SOD1^D90D SKM), and 34 (SOD1^D90D SKM) ROIs were analyzed, with each dot representing one ROI. All data were collected from three independent experimental replicates. The unpaired 2-tailed Student’s t-test was used in (B–F) and one-way analysis of variance with Tukey’s post hoc test was used in (H). Scale bars: 20 μm in confocal 3D images; and 5 μm in 2D image slices.