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Ce-DAB2 staining and preparation of Drosophila larval CNS for EM The dissected flies’ central brain and VNC were rinsed (five times for 3 min each) with 0.15 M sodium cacodylate buffer pH 7.4 containing 2 mM CaCl 2 on ice and blocked with 20 mM glycine, 50 mM potassium cyanide and 50 mM aminotriazole in 0.15 M sodium cacodylate buffer pH 7.4 containing 2 mM CaCl 2 for 20 min on ice

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Proteoglycans like hyaluronan, decorin, versican, and perlecan bind water molecules and growth factors, influencing extracellular hydration and biomechanical characteristics [9]