Journal: Aging Cell
Article Title: Synaptic Vesicle Glycoprotein 2A Suppresses Amyloidogenesis Beyond Its Synaptic Role: A Novel Mechanism Disrupting BACE1 Binding and Altering APP Localization
doi: 10.1111/acel.70379
Figure Lengend Snippet: Effect of SV2A overexpression on the distribution of APP. (a, b) Expressions of EEA1, Rab7, Rab11, and LAMP1 in SV2A‐overexpressed SH‐SY5Y cells (N2a cells) and their control cells, as well as the relative expression analysis. (c, d) Fluorescence intensity of EEA1 and APP in SV2A‐overexpressed SH‐SY5Y cells (N2a cells) and the co‐localization analysis of EEA1 and APP. (e, f) Fluorescence intensities of Rab7 and APP in SV2A‐overexpressed SH‐SY5Y cells (N2a cells) and the co‐localization analysis of Rab7 and APP. (g, h) Fluorescence intensity of Rab11 and APP in SV2A‐overexpressed SH‐SY5Y cells (N2a cells) and the co‐localization analysis of Rab11 and APP. (i–j) Fluorescence intensities of LAMP1 and APP in SV2A‐overexpressed SH‐SY5Y cells (N2a cells) and the co‐localization analysis of LAMP1 and APP. (k) Quantification of surface APP expression in SH‐SY5Y cells transfected with SV2Aoe or control plasmids. (l) Quantification of surface APP fluorescence intensity (APP/DAPI) in SH‐SY5Y cells transfected with SV2Aoe or control plasmids, scale bar: 10 μm. (m) Fluorescence level of APP on the cell surface in hippocampal regions of APP/PS1 mice injected with AAV‐SV2A or AAV‐Con; scale bar: 50 μm. (n) Relative fluorescence intensity of APP (APP/DAPI) on the cell surface in the hippocampal regions of APP/PS1 mice injected with AAV‐SV2A or AAV‐Con. (o) Fluorescence level of APP on the cell surface in cortical regions of APP/PS1 mice injected with AAV‐SV2A or AAV‐Con; scale bar: 50 μm. (p) Relative fluorescence intensity of APP (APP/DAPI) on the cell surface in the cortical regions of APP/PS1 mice injected with AAV‐SV2A or AAV‐Con. The protein levels of APP on the cell surface were measured by the cell surface biotin assay. Data are presented as the mean ± SD. All dot plots: t ‐test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.
Article Snippet: The following primary antibodies were used in the study, early endosome antigen 1 (EEA1) (Cat. No. sc‐137,130, Santa Cruz, 1:100); APP (Cat. No. ab32136, Abcam, 1:2000; Cat. No. MAB348, Sigma‐Aldrich, 1:400); BACE1 (Cat. No. PA1‐757, Thermo Fisher Scientific, 1:200); Ras‐related protein Rab‐7 (Rab7) (ab137029, Abcam, 1:100); Ras‐related protein Rab‐11 (Rab11) (Cat. No. 5589, Cell Signaling Technology, 1:100); Lysosome‐associated membrane protein‐1 (LAMP1) (Cat. No. 99437, Cell Signaling Technology, 1:100).
Techniques: Over Expression, Control, Expressing, Fluorescence, Transfection, Injection
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