Fig. 6.

UA induces the expression of autophagy-related genes, increases autophagy flux and clears Aβ in vitro. A Relative expression of autophagy-related genes assessed via qPCR from mRNA extracted from SH-SY55 (human neuroblastoma—right side) and HT-22 (mouse hippocampal—left side) cells treated with 30 μM UA versus vehicle-treated controls (DMSO) for 24 h. Data is presented as mean fold change in relative units (ru) ± SEM by normalizing expression of each gene to that of human β-actin (ACTB) or mouse glyceraldehyde-3-phosphate dehydrogenase (Gapdh). *p value < 0.05 calculated via unpaired t test. B Representative single-plane confocal micrographs of HT-22 cells treated with or without UA (30 μM) and/or bafilomycin (50 nM) showing fluorescence of DQ-BSA red (red signal) and DAPI nuclear staining (blue signal) after 6 h of uptake and respective drug treatments. Scale bars = 30 μm. Corresponding pixel quantification data is presented as mean fold change in relative units (ru) ± SEM; p value was calculated via unpaired t test, n = 3. C Western blot analysis of amyloid beta, SQSTM1, and LC3B protein levels, using specific antibodies in HT-22 cells treated with Aβ oligomers (2 μM, 48 h), with or without UA and/or bafilomycin. Shown is a representative figure for four independent experiments. Quantification of changes in protein expression is presented as mean fold change in relative units (ru) ± SEM by normalizing each protein’s band intensity to the expression of actin and are presented as a fold-change relative to the samples treated with DMSO as vehicle. *p value < 0.05 calculated via unpaired t test