rabbit anti rab11  (Cell Signaling Technology Inc)

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: bioRxiv

Article Title: FAM84B facilitates tau propagation via RYR3-mediated exocytosis in response to neuroinflammation

doi: 10.64898/2025.12.28.696549

Figure Lengend Snippet: (A) Levels of caveolin-1, CHC, EEA1, RAB5, RAB7, RAB9, RAB11, and actin in SH-SY5Y cells expressing tau alone or tau with FAM84B-Myc (n=3 each). (B) CHC levels in the cerebral cortex of HC (n=3) and patients with VD (n=3), CBD (n=3), PSP (n=4), PART (n=3), and AD (n=6). (C) Levels of CHC and actin in iPSC-derived CN from HC and patients with AD (n=3 each). (D) Changes in the levels of tau and FAM84B (Myc) in the CM from SH-SY5Y cells expressing tau alone or tau with FAM84B-Myc after treatment with either DMSO or Pitstop2 (n=3 each). (E) Changes in the levels of phosphorylated tau (S396) in the CM from SH-SY5Y cells expressing tau alone or tau with FAM84B-Myc after treatment with either DMSO or Pitstop2 (n=8 each). (F) Changes in the levels of PHF-1, total tau, FAM84B (Myc), CHC, and actin in SH-SY5Y cells expressing tau alone or tau with FAM84B-Myc after treatment with either DMSO or Pitstop2 (n=8 each). (G) PHF-1 spread from SH-SY5Y cells expressing Tau-p2a-eGFP with FAM84B to neighboring cells in response to treatment with either DMSO or Pitstop2 (n=4 each). Arrows indicate recipient cells (red) to which tau has been transmitted. Solid boxes in the images indicate magnified areas. Results are presented as mean ± SEM. Statistical analyses: (B) ordinary two-way ANOVA; (A, C-G) unpaired two-tailed t -test with Welch’s correction; ns: not significant, *p < 0.05, **p < 0.01, ***p < 0.001. Abbreviations: CHC, clathrin heavy chain; HC, healthy control; VD, vascular dementia; CBD, corticobasal degeneration; PSP, progressive supranuclear palsy; PART, primary age-related tauopathy; AD, Alzheimer’s disease; CN, cortical neuron; CM, conditioned medium; PHF-1, phosphorylated tau at S396/S404 residue; FOV, field of view.

Article Snippet: Following blocking, membranes were incubated overnight at 4°C with primary antibodies (1:1,000 dilution), including: PHF-1 (Peter Davies Lab), Tau5 (Gail VW Johnson Lab), Tau (Dako, Agilent Technologies, Inc., 0024), Myc-tag (Cell Signaling Technology, 2276), FAM84B (OriGene, TA501992; Proteintech Group, Inc., Rosemont, IL, USA, 18421-1-AP), Alix (Cell Signaling Technology, 2171), Caveolin-1 (Cell Signaling Technology, 3267), RYR3 (Alomone Labs, Jerusalem, Israel, ARR003), Clathrin-heavy chain (CHC, Cell Signaling Technology, 4796), EEA1 (Cell Signaling Technology, 3288), Rab5 (Cell Signaling Technology, 3547), Rab7 (Cell Signaling Technology, 9367), Rab9 (Cell Signaling Technology, 5118), Rab11 (Cell Signaling Technology, 5589), pSTAT3 (Y705) (Cell Signaling Technology, 9145), STAT3 (Cell Signaling Technology, 9139), p62 (Cell Signaling Technology, 8025), LC3 (Cell Signaling Technology, 3868), HSP70 (Cell Signaling Technology, 4876), LAMP-1 (BD Biosciences, Franklin Lakes, NJ, USA, 611042), LAMP-2 (Sigma-Aldrich, L0668), and beta-actin (Merck Millipore, MAB1501).

Techniques: Expressing, Derivative Assay, Two Tailed Test, Control, Residue