Journal: bioRxiv

Article Title: Isolation and characterization of synaptic structures from human neural organoids

doi: 10.1101/2025.01.21.634055

Figure Lengend Snippet: A) Changes in the levels of 30 selected synaptic markers in the differential centrifugation fractions compared to the homogenate from each tissue type. An enrichment of synaptosomes from adult mouse brain (W8) using the conventional Percoll® density gradient centrifugation method was performed in parallel to compare the synaptic protein enrichment levels. The conventional Percoll® F3 and F4 synaptosome enriched fractions are shown. **p < 0.001; ***p < 0.0001. B) Heatmaps of the normalized abundances of the same 30 synaptic markers as used in A for each fraction and tissue or enrichment type. P1: pellet from the first centrifugation, containing mainly whole cells, cell debris and nuclei to serve as a negative control. C) Western blots of the synaptic marker SYP in differential centrifugation fractions from human forebrain organoids and human cortical tissue.

Article Snippet: Carefully, 2 mL of S1 was layered on top of each Percoll density gradient (4 layers of 3%, 10%, 15% and 23% Percoll (Cytiva) in sucrose/EDTA buffer with 0.25 mM DTT) and centrifuged at 20,000 rpm for 5 min when reaching top speed in a Sorval RC 5C plus Harvest centrifuge using a Sorval SS34 rotor.

Techniques: Centrifugation, Gradient Centrifugation, Protein Enrichment, Negative Control, Western Blot, Marker