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

Journal: Drug Delivery

Article Title: Cell swelling and upright mounting-based imaging for high-resolution visualization of intracellular trafficking across the BBB using conventional confocal microscopy

doi: 10.1080/10717544.2025.2608235

Figure Lengend Snippet: Co-localization analysis of A647-Tf (A) and A647-anti-TfR Ab (B) with intracellular markers over time. Manders’ overlap coefficients were calculated from multiple xy sections within z-stack confocal images acquired using NIS-Elements (NIS-E) software. Cells were treated with A647-Tf or A647-anti-TfR Ab (5 μg/mL) for 5 min, 15 min, or 1 h, and stained with markers for early endosomes (EE: Rab5, EEA1), late endosomes (LE: Rab7, VAMP7), lysosomes (Ly: LAMP1), recycling endosomes (RE: Rab11, VAMP3), and basolateral membrane (BM: SNAP23, syntaxin 4). Data are presented as mean ± SEM ( n = 6, each image was obtained from one sample in a single experiment).

Article Snippet: Early Endosome Antigen 1 (EEA1) , Early Endosome (EE) , 1:1000 , Cell Signaling Technology , 3288S.

Techniques: Software, Staining, Membrane

Journal: Drug Delivery

Article Title: Cell swelling and upright mounting-based imaging for high-resolution visualization of intracellular trafficking across the BBB using conventional confocal microscopy

doi: 10.1080/10717544.2025.2608235

Figure Lengend Snippet: Confocal microscopy analysis of early endosome trafficking patterns. (A) Cells treated with A647-Tf for 5 min, 15 min, or 1 h, stained with DAPI (blue), Rab5 (green, upper panels), and EEA1 (green, lower panels). (B) Cells treated with A647-anti-TfR Ab under the same conditions and staining. The red signal corresponds to internalized A647-Tf or A647-anti-TfR Ab. Scale bar: 10 μm.

Article Snippet: Early Endosome Antigen 1 (EEA1) , Early Endosome (EE) , 1:1000 , Cell Signaling Technology , 3288S.

Techniques: Confocal Microscopy, Staining