Journal: bioRxiv

Article Title: NATO3 protects dopaminergic neurons in mouse in vivo and human in vitro Parkinson’s disease models

doi: 10.1101/2025.10.10.681573

Figure Lengend Snippet: a. Experimental timeline. b. Schematic of the lentiviral vector for bicistronic expression of NATO3 and EGFP from the PGK promoter, and the control vector expressing only EGFP. c. Representative images of iPSC-derived DA neurons immunostained for α-Syn, TH, and GFP. PD, cells differentiated from an iPSC line carrying the SNCA A53T mutation. Control, mutation-corrected isogenic control cells. NATO3, cells transduced with the NATO3 vector. GFP, cells transduced with the control vector. Scale bar, 50 µm. d. Quantification of α-Syn fluorescence intensity within DA neurons at DIV90. e. Representative images of iPSC-derived DA neurons stained with Amytracker 630 (AmyT), co-stained for TH and GFP. Scale bar, 20 µm. f. Quantification of Amytracker fluorescence intensity within DA neurons at DIV90. In d and f, scatter plots indicate mean ± SD. Each data point represents the mean fluorescence intensity value from a single TH-positive cell. All data are from N=3 independent differentiation experiments. Statistical comparisons were performed using a Kruskal-Wallis test with a Dunn’s multiple comparisons test. *p<0.05, **p<0.01, ***p<0.001, and ****p<0.0001. ns, not significant.

Article Snippet: This synthetic construct was custom-made by Eurofins and cloned into a lentiviral transfer plasmid backbone.

Techniques: Plasmid Preparation, Expressing, Control, Derivative Assay, Mutagenesis, Transduction, Fluorescence, Staining