third generation lentiviral vector pcw57 1  (Addgene inc)

Journal: Antioxidants

Article Title: Exo70 Protects Against Memory and Synaptic Impairments Following Mild Traumatic Brain Injury

doi: 10.3390/antiox14060640

Figure Lengend Snippet: Exo70 overexpression reinforces NMDAR synaptic availability upon mTBI induction. Samples from the dorsal hippocampus were analyzed by Western blot. Here, 30 µg of protein samples were resolved in 10% SDS-PAGE and transferred to PVDF membranes. Tubulin and Actin were used as loading controls. ( A ) Characterization of the lentiviral expression in all experimental groups using GFP and HA antibodies. ( B ) Densitometric analysis of HA-Exo70. ( C ) Western blot analyzing NMDAR synaptic localization after mTBI in injected mice. Samples were analyzed using GluN2B p1472, GluN2B p1336, and total GluN2B antibodies. ( D – F ) Densitometric analysis normalized with loading controls before normalization with Sham-GFP control samples. ( G ) Western blot analyzing NMDAR synaptic-signaling-related proteins after mTBI induction in Exo70-overexpressing samples. Samples were analyzed using pCREB and pERK1/2 antibodies. ( H , I ) The graphs show the densitometric analysis normalized with loading controls before normalization with Sham-GFP control samples. White circles indicate individual experiments. Values represent means ± SEM, n = 5 mice per experimental group. Statistical differences were calculated by two-way ANOVA, followed by post hoc Bonferroni’s test. * p < 0.05, ** p < 0.01, *** p < 0.001, ns: statistically non-significant difference.

Article Snippet: Third-generation lentiviral particles were packaged by and acquired from Applied Biological Materials (ABM, Canada) based on the lentiviral vector FUG-1D2A-Exo70-W [ ].

Techniques: Over Expression, Western Blot, SDS Page, Expressing, Injection, Control