Fig. 2

Synaptic stimulation increases peripheral CSP density in Atl-KD larvae A Illustration showing the protocol followed for the acquisition of NMJs STED images, with (S) or without (US) KCl Stimulation; larvae were fixed and processed for immunostaining after the protocol. Quantification of the CSP particle density was performed in the peripheral delimited area (ROI generated from the external limit of HRP staining to 200 µm inside the bouton), as well as in the center of the bouton. Cartoon created with BioRender.com. B Normalized CSP intensity in larval boutons of different genotypes. Each dot represents the average density of at least 10 boutons in one larva. ** = Mann-Whitney p-value < 0.01. n = 7-9 larvae. C–D Representative images of CSP staining in synaptic boutons of Atl-KD and Tkv-CA larvae. Scale bar: 2 µm. E–F CSP density in boutons of unstimulated (US) and KCl-stimulated (S) NMJs. Peripheral E and central F CSP density of Atl-KD and Tkv-CA larvae. G–H Vesicle mobilization (CSP density in S boutons)/(CSP density in US boutons) in Atl-KD and Tkv-CA larvae, in the periphery G and the center of the bouton H each data point represents one bouton from 5 different larvae. I–J CSP density distribution on the whole synaptic bouton by 10% segment bands, under non-stimulated (US) and stimulated (S) conditions, for the different genotypes. Each data point represents the average of all boutons with CSP particles in that segment band. One-way ANOVA, p-value * < 0,05; ** < 0.01; *** < 0.001; **** < 0.0001. n = 5 larvae