Fig. 3
From: Metalloprotease Adam10 suppresses epilepsy through repression of hippocampal neuroinflammation
AAV-Adam10 vector construction and hippocampal Adam10 overexpression verification. a Structure of pHBAAV-CAG-MCS-T2A-ZsGreen AAV vector, which carries Adam10 and an empty construct or a ZsGreen gene as an indicator. b Single-clone PCR identification of Adam10 expression (lanes 1–5). c Graphic illustration of the AAV bilateral injection sites in the hippocampus of the mouse brain. Arrows indicate the bilateral injection sites in the hippocampal CA1 region. d Distribution of AAV-mediated ZsGreen expression in the CA1 region of the hippocampus. e, f RT-PCR analysis of Adam10 expression in the hippocampus of Vehicle Ctrl, AAV-Ctrl, and AAV-Adam10 mice, respectively (F2,12 = 13.41, p = 0.007, AAV-Adam10 vs Vehicle Ctrl; p = 0.004, AAV-Adam10 vs AAV-Ctrl) (n = 5). g, h Western blot analysis of Adam10 protein levels in the hippocampus of Vehicle Ctrl, AAV-Ctrl, and AAV-Adam10 mice, respectively (F2,12 = 12.06, p = 0.002, AAV-Adam10 vs Vehicle Ctrl; p = 0.004, AAV-Adam10 vs AAV-Ctrl) (n = 5). i Representative images of the Adam10 immunostaining in the hippocampal CA1 region of AAV-Ctrl and AAV-Adam10 mice, respectively. j Bar graphs showing the quantification of Adam10-positive cells in the hippocampal CA1 region of the AAV-Ctrl and AAV-Adam10 mice, respectively (p = 0.012) (n = 4). *p < 0.05, **p < 0.01, unpaired two-tailed Student’s t test, and one-way ANOVA. Scale bar = 50 μm in d and 20 μm in i