Journal: Science Advances
Article Title: Lipid nanoparticles for mRNA delivery in brain via systemic administration
doi: 10.1126/sciadv.adw0730
Figure Lengend Snippet: Flow cytometry analysis of GFP expression ( A ) or tdTomato expression ( B ) across different brain cells [neuronal nuclei (NeuN) + for neuron, glial fibrillary acidic protein (GFAP) + for astrocyte, CD11b + for microglia, and CD31 + for BCEC]. Data are presented as means ± SD in (A) and (C) ( n = 3 biological replicates). Statistical significance was analyzed by one-way ANOVA with Tukey’s multiple comparisons test. ( C ) Schematic illustrating the delivery of Cre mRNA to activate tdTomato expression, along with the administration regimen. ( D ) Representative immunofluorescence sections of brain tissues of Ai14 mice after three administrations with PBS, MC3, or OS4T LNPs (Cre mRNA, 1 mg/kg). Scale bar, 2.5 mm. The tdTomato expression in neurons ( E ), microglia ( F ), and astrocytes ( G ) after three administrations with PBS, MC3, or OS4T LNPs (Cre mRNA, 1 mg/kg). Scale bar, 50 μm. ** P < 0.01; *** P < 0.001; **** P < 0.0001.
Article Snippet: The following antibodies were used: NeuN antibody (1:50 dilution, Miltenyi Biotec, clone: REA1131) for neurons; GFAP antibody (1:300 dilution, Cell Signaling Technology, cat. no. 3655S) for astrocytes; anti-Iba1 antibody (1:5000 dilution, Abcam, ab283346) and goat anti-rat IgG H&L [fluorescein isothiocyanate (FITC)] (1:1000 dilution, Abcam, ab6840) for microglia; and tdTomato (E3G5L) rabbit monoclonal antibody (1:300 dilution, Cell Signaling Technology, cat. no. 20163) and goat anti-rabbit IgG H&L [Cyanine3 (Cy3)] (1:100 dilution, Abcam, ab6939) for tdTomato.
Techniques: Flow Cytometry, Expressing, Immunofluorescence