Journal: Nature Aging

Article Title: The VCAM1–ApoE pathway directs microglial chemotaxis and alleviates Alzheimer’s disease pathology

doi: 10.1038/s43587-023-00491-1

Figure Lengend Snippet: a , STRINGdb protein–protein interaction between VCAM1 (green) and Aβ plaque-associated proteins (orange). b , Schematic diagram showing the protocol for injecting protein-coated beads followed by IL-33 treatment in APP/PS1 mice. c– e , Representative images (c) and bar plots showing the numbers of microglia (d) and MHC-II + microglia (e) within the area of BSA-, ApoE-, CD44-, and ITGB2-coated beads after IL-33 treatment (Con: BSA: n = 3, ApoE: n = 6, CD44: n = 6, ITGB2: n = 4; IL-33: BSA: n = 5, ApoE: n = 6, CD44: n = 6, ITGB2: n = 5; two-way ANOVA with Šidák’s multiple comparisons test). Dotted line indicates bead area. Scale bar = 20 μm. f , Representative images showing the Vcam1 -expressing microglia surrounding ApoE-coated beads after IL-33 treatment. Arrowheads indicate Vcam1 -expressing microglia. Scale bars = 50 μm (left) and 20 μm (right). Experiment was repeated for three batches with similar results. g –i, Representative images (g) and bar plots showing the numbers of microglia (h) and MHC-II + microglia (i) within the areas of beads coated with nonlipidated or lipidated ApoE in IL-33-treated APP/PS1 mice (nonlipidated: n = 7; lipidated: n = 7; two-tailed paired t -test). Dotted line indicates bead area. Scale bar = 20 μm. j–o , VCAM1 is essential for the ApoE chemotaxis of microglia after IL-33 treatment. j– l , Representative images (j) and bar plots showing the numbers of microglia (k) and MHC-II + microglia (l) within the ApoE-coated bead areas after co-injection of rVCAM1 in IL-33-treated APP/PS1 mice (Fc: n = 6; rVCAM1: n = 6; two-tailed paired t -test). Dotted line indicates bead area. Scale bar = 20 μm. m – o , Representative images (m) and bar plots showing the numbers of microglia (n) and MHC-II + microglia (o) within the ApoE-coated bead areas in IL-33-treated APP/PS1;VCAM1-icKO mice (wild-type [WT]: n = 6; icKO: n = 6; two-tailed unpaired t -test). Dotted line indicates bead area. Scale bar = 20 μm. All data are mean ± s.e.m.

Article Snippet: We obtained mouse ApoE recombinant protein (MBS955382) from MyBioSource as well as CCR7 neutralizing (clone: 4B12) antibody (MAB3477) and VCAM1 antibody (BBA5) from R&D Systems.

Techniques: Expressing, Two Tailed Test, Chemotaxis Assay, Injection

Journal: Nature Aging

Article Title: The VCAM1–ApoE pathway directs microglial chemotaxis and alleviates Alzheimer’s disease pathology

doi: 10.1038/s43587-023-00491-1

Figure Lengend Snippet: a , b , Quality control for bead injection. Representative images (a) and quantification (b) showing the bead injection area (Con: n = 12; interleukin-33 [IL-33]: n = 18, two-tailed unpaired Student’s t -test). Scale bar = 50 μm. c , d , IL-33-induced chemotactic microglia migrate towards human ApoE isoforms. Representative images (c) and bar plot (d) showing microglial migration towards BSA-, murine ApoE (mApoE)-, human ApoE3-, and human ApoE4-coated beads after IL-33 treatment ( n = 3 per condition; one-way ANOVA with Šidák’s multiple comparisons test). Dotted line indicates bead area. Scale bar = 25 μm. All data are mean ± s.e.m.

Article Snippet: We obtained mouse ApoE recombinant protein (MBS955382) from MyBioSource as well as CCR7 neutralizing (clone: 4B12) antibody (MAB3477) and VCAM1 antibody (BBA5) from R&D Systems.

Techniques: Control, Injection, Two Tailed Test, Migration

Journal: Nature Aging

Article Title: The VCAM1–ApoE pathway directs microglial chemotaxis and alleviates Alzheimer’s disease pathology

doi: 10.1038/s43587-023-00491-1

Figure Lengend Snippet: a–c , ApoE-neutralizing antibody inhibits the Aβ chemotaxis of microglia upon IL-33 treatment. a , Schematic diagram showing the protocol for ApoE-neutralizing antibody administration before IL-33 treatment in APP/PS1 mice. b , c , Representative images (b) and violin plot (c) showing the distance between chemotactic microglia (that is, Vcam1 + Cx3cr1 + cells) and the nearest Aβ plaque 24 h after administration of ApoE-neutralizing antibody in IL-33-treated APP/PS1 mice (IgG Con: n = 107 microglia from 6 mice; IgG IL-33: n = 115 microglia from 5 mice; αApoE Con: n = 94 microglia from 6 mice; αApoE IL-33: n = 93 microglia from 6 mice; Kruskal–Wallis test with Dunn’s multiple comparisons test). Dotted circle indicates 10 μm from the perimeter of the Aβ plaque. Arrowheads indicate Vcam1 -expressing microglia. Scale bar = 10 μm. d – i , VCAM1–ApoE interaction is required for inducing the phagocytic state transition of microglia after the induction of VCAM1 expression. d – f , Representative images (d) and bar plots showing the proportions of Aβ plaque-associated microglia (e) and phagocytic microglia (f) (that is, MHC-II + Iba1 + cells) 24 h after administration of ApoE-neutralizing antibody in IL-33-treated APP/PS1 mice (IgG Con: n = 5; IgG IL-33: n = 5; αApoE Con: n = 5; αApoE IL-33: n = 5; two-way ANOVA with Šidák’s multiple comparisons test). Arrowheads indicate phagocytic microglia. Scale bar = 20 μm. g – i , Representative images (g) and bar plots showing the proportions of Aβ plaque-associated microglia (h) and phagocytic microglia (i) (that is, MHC-II + Iba1 + cells) 24 h after IL-33 treatment in APP/PS1-ApoE–knockout mice ( n = 6 per condition; two-way ANOVA with Šidák’s multiple comparisons test). Scale bar = 20 μm. j , k , Representative images (j) and bar plot (k) showing the Aβ plaque areas in the cortex 48 h after administration of ApoE-neutralizing antibody in IL-33-treated APP/PS1 mice (IgG Con: n = 5; IgG IL-33: n = 7; αApoE Con: n = 4; αApoE IL-33: n = 4; two-way ANOVA with Šidák’s multiple comparisons test). Scale bar = 200 μm. All data are mean ± s.e.m.

Article Snippet: We obtained mouse ApoE recombinant protein (MBS955382) from MyBioSource as well as CCR7 neutralizing (clone: 4B12) antibody (MAB3477) and VCAM1 antibody (BBA5) from R&D Systems.

Techniques: Chemotaxis Assay, Expressing, Knock-Out

Journal: Journal of Neuroinflammation

Article Title: The protective PLCγ2-P522R variant mitigates Alzheimer’s disease-associated pathologies by enhancing beneficial microglial functions

doi: 10.1186/s12974-025-03387-6

Figure Lengend Snippet: PLCγ2-P522R variant does not increase the plaque-associated APOE or DAM signature in female APP/PS1 mice. A Analysis of total APOE and APOE areas (µm 2 ) within and surrounding (within 0–10- and 10–20-µm distances from the amyloid plaque outline) β-amyloid plaques revealed no differences between the APP/PS1xPLCγ2-P522R (A+/P ki/ki ) and APP/PS1 (A+/P wt/wt ) mice. A+/P wt/wt n = 5, A+/P ki/ki n = 6. Colored disks on the x-axis indicate the analyzed area overlapping (middle circle) and surrounding β-amyloid plaques. The scale bar in the representative immunofluorescence images is 50 μm. Unpaired samples t test. All the data are presented as the means ± SEMs. Each data point represents an individual mouse. B Heatmap of z scored vst-normalized expression of homeostatic as well as stage 1 and stage 2 disease-associated microglia (DAM) signature genes in the bulk temporo-occipital cortex and C ) CD11b+ microglia isolated from the whole brain tissue of 13-month-old wild-type (A-/P wt/wt ), A+/P wt/wt and A+/P ki/ki female mice. Cortex A-/P wt/wt n = 1, A+/P wt/wt n = 4, and A+/P ki/ki n = 4; CD11b+ A-/P wt/wt n = 7, A+/P wt/wt n = 4, and A+/P. ki/ki n = 4

Article Snippet: Soluble and insoluble APOE were measured in lysates using an Apolipoprotein E (APOE) Mouse ELISA Kit (#ELK2007, Gentaur) according to the manufacturer’s instructions.

Techniques: Variant Assay, Immunofluorescence, Expressing, Isolation

Journal: iScience

Article Title: INPP5D modulates TREM2 loss-of-function phenotypes in a β-amyloidosis mouse model

doi: 10.1016/j.isci.2023.106375

Figure Lengend Snippet: Key resources table

Article Snippet: Goat polyclonal anti-Apolipoprotein E , Merck/Millipore , Cat# AB947, RRID: AB_2258475.

Techniques: Plasmid Preparation, Purification, Control, Recombinant, Transfection, Enzyme-linked Immunosorbent Assay, Blocking Assay, Bicinchoninic Acid Protein Assay, Cell Culture, Membrane, Pore Size, In Situ, Labeling, Isolation, Software