Fig. 5

Synapsin-1 localization at the adult NMJ. A Western blot analysis and data quantification of diaphragm samples showing specificity of the anti-SNAP25/pSNAP25 and anti-Synapsin-1/pSynapsin-1 antibodies to show protein bands appear in the correct molecular weight. B–E1 Neuromuscular junctions of levator auris longus (LAL) muscle visualized at confocal microscopy. B Confocal images of a NMJ with triple labeling: SNAP-25 (green), Syntaxin (blue), and AChR (red). Scale bar, 10 μm. C Confocal images of multiple NMJ stained with Synapsin-1 (green) and AChR (red). Scale bars, 50 μm. D Confocal images of an NMJ with triple labeling: Synapsin-1 (green), Syntaxin (red), and AChR (blue). Scale bar, 10 μm. D1–D3 Confocal optical section of the NMJ shown in B. Scale bars, 10 μm. “#” shows areas of the nerve terminal rich in Synapsin-1 and poor in Syntaxin. “*” shows areas of the nerve terminal rich in Syntaxin and poor in Synapsin-1. E Confocal images of a NMJ with triple labeling: Synapsin-1 (green), Syntaxin (red), and AChR (blue). Scale bar, 10 μm. E1 Scale bar, 3 μm. Arrows show areas of the nerve terminal rich in Syntaxin and poor in Synapsin-1. F Quantitative colocalization represented as heatmap of the intensity of between anti-Synapsin-1 labeling, presynaptic anti-Syntaxin labeling and postsynaptic AChR labeling. F1 Average Pearson’s correlation coefficient of the colocalization between Synapsin-1 versus Syntaxin or Synapsin-1 versus AChRs. ***p < 0.001 versus the corresponding comparison