ebss (earle's balanced salt solution)  (HiMedia Laboratories)

Journal: Neural Regeneration Research

Article Title: Enhanced autophagic clearance of amyloid-β via histone deacetylase 6-mediated V-ATPase assembly and lysosomal acidification protects against Alzheimer’s disease in vitro and in vivo

doi: 10.4103/NRR.NRR-D-23-01633

Figure Lengend Snippet: VPA activates autophagy and promotes lysosomal function in an in vitro model of Alzheimer’s disease. (A) Detection of mRFP-GFP-LC3 delivery to lysosomes. SH-SY5Y cells (WT), VEH-treated APP swe cells, and VPA-treated APP swe cells were treated with EBSS, and mRFP-GFP-LC3 fluorescent signals were detected by confocal microscopy. Scale bar: 5 μm. (B, C) mRFP-positive and GFP-positive puncta were quantified to determine the number of autophagosomes and autolysosomes per cell ( n = 6 randomly selected fields). (D) Detection of mRFP-GFP-LC3 delivery to lysosomes. SH-SY5Y cells (WT), VEH-treated APP swe cells, and VPA-treated APP swe cells were treated with Baf A1. Scale bar: 10 μm. (E, F) mRFP and GFP puncta were quantified to determine the number of autophagosomes and autolysosomes per cell ( n = 6 randomly selected fields). * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way analysis of variance followed by Tukey’s post hoc test). Baf A1: Bafilomycin A1; EBSS: Earle’s Balanced Salt Solution; mRFP-GFP-LC3: monomeric red fluorescence protein–enhanced green fluorescence protein-microtubule-associated protein 1 light chain3; VEH: vehicle; VPA: valproic acid.

Article Snippet: Then, the cells were treated with Earle’s Balanced Salt Solution (EBSS) (Thermo Fisher Scientific) for 4 hours to induce autophagy or Bafilomycin A1 (Baf A1, MedChem Express, Monmouth Junction, NJ, USA) for 4 hours to inhibit autophagy.

Techniques: In Vitro, Confocal Microscopy, Fluorescence