antibody anti hvcn1  (Alomone Labs)

Journal: Experimental & Molecular Medicine

Article Title: Hv1 inhibition rescues AD pathology by restoring microglial mitochondrial function and enhancing mitochondrial transfer

doi: 10.1038/s12276-025-01593-z

Figure Lengend Snippet: a A schematic representation of the intrahippocampal injection protocol. The virus constructs AAV–EGFP and AAV-hTAU were infused into the hippocampus of 2-month-old Hvcn1 −/− mice and littermate controls. Mice were killed 40 days post-injection for immunofluorescence analysis. b Representative images of Iba1 staining and 3D skeletonization in the hippocampus of Hvcn1 −/− mice and their littermates injected with AAV–EGFP ( Hvcn1 −/− + AAV–EGFP and WT + AAV–EGFP) or AAV-hTAU ( Hvcn1 −/− + AAV-hTAU and WT + AAV-hTAU). The red dots indicate branching points, and the green dots indicate terminal points. c A quantification of microglial branch number, terminal points and filament length using Imaris software ( n = 16-27 cells from three to five mice). d A quantification of the number of Iba1 + microglia per square millimeter in the hippocampus ( n = 14-23 fields of view from three to five mice). e , f Representative immunofluorescent images and corresponding 3D skeletonization of microglia ( e ) and quantification of branch number, terminal points and filament length ( f ) in the hippocampus of C57BL/6 mice injected with AAV–EGFP (AAV-CTRL), AAV-hTAU (AAV-TAU) or AAV-hTAU with YHV98-4 treatment (AAV-TAU + YHV98-4) ( n = 16–19 cells from five mice). g A quantification of the number of Iba1 + microglia per square millimeter in the hippocampus ( n = 22 fields of view from five mice). h , i Representative immunofluorescent images showing the colocalization of DAPI (blue), Hv1 (green) with Iba1 (red), NeuN (red) or GFAP (red) ( h ) and quantification of Hv1 intensity ( i ) ( n = 19 fields of view from five mice). j Hvcn1 expression in different microglial subclusters from patients with AD with varying Braak stages. The data were calculated using a one-way ANOVA followed by a Tukey’s post hoc analysis and presented as the mean ± s.e.m. The P values are presented on the graph.

Article Snippet: The primary antibodies used were: anti-Iba1 (1:100, ab283346, Abcam), anti-Iba1 (1:200, A19776, ABclonal), anti-NeuN (1:500, ab279296, Abcam), anti-NeuN (1:400, ab104224, Abcam), anti-MAP2 (1:200, 8707, Cell Signaling Technology), anti-AT8 (phospho-Tau (S202/T205), 1:600, GB113883 , Servicebio), anti-CD44 (1:500, GB112054 , Servicebio), anti-Hv1(1:100, AHC-001, Alomone Labs), anti-CD11c (1:100, ab254183, Abcam), anti-mouse I-A/I-E (1:100, 107601, BioLegend), anti-nicotinamide adenine dinucleotide phosphate oxidase 2 (NOX2) (1:200, GB11391-100, Servicebio), anti-GFAP (1:500, BD12096, Servicebio), anti-CD4 (1:100, 100505, BioLegend) and anti-CD8 (1:200, GB114196 , Servicebio) antibodies.

Techniques: Injection, Virus, Construct, Immunofluorescence, Staining, Software, Expressing

Journal: Experimental & Molecular Medicine

Article Title: Hv1 inhibition rescues AD pathology by restoring microglial mitochondrial function and enhancing mitochondrial transfer

doi: 10.1038/s12276-025-01593-z

Figure Lengend Snippet: a–c Representative staining of Iba1 + microglia and corresponding 3D skeletonization ( a ), quantification of microglial morphology ( b ) ( n = 18–23 cells from five mice) and microglial number ( c ) ( n = 20–23 fields of view from five mice) in WT, 3×Tg and 3×Tg+YHV98-4 mice. d , e Representative images showing Hv1 (green) costained with Iba1 (red) and DAPI (blue) ( d ) and quantification of Hv1 intensity ( e ) in the hippocampus ( n = 20 fields of view from five mice). f , g Representative images of 4G8 staining in the hippocampus ( f ) and quantification of 4G8 + area ( g ) ( n = 3 mice). h , i Representative AT8 staining ( h ) and quantification of AT8 coverage and intensity ( i ) in the hippocampus ( n = 21–24 fields of view from five mice). j , k Representative images of DHE staining in the hippocampus ( j ) and quantification of DHE fluorescence intensity ( k ) ( n = 18 fields of view from three mice). l GSEA of significant downregulated pathways in microglia of 3×Tg versus WT mice. m GSEA of significantly upregulated pathways in microglia of 3×Tg+YHV98-4 versus 3×Tg mice. n – p Representative images of NeuN and MAP2 staining ( n ) and quantification of NeuN intensity ( o ) and MAP2 coverage ( p ) ( n = 19–20 fields of view from five mice). q – v Behavioral assessments in WT ( n = 10), 3×Tg ( n = 8) and 3×Tg+YHV98-4 mice ( n = 11) mice. Escape latency during MWM training trials ( q ), quantification of platform crossings ( r ) and time spent in the target quadrant ( s ), and representative track paths during the probe trials ( t ). A quantification of the discrimination index in the NOR test ( u ) and the percentage of spontaneous alternation in the Y-maze test ( v ). The data were calculated using a one-way ANOVA followed by a Tukey’s post hoc test in b , c , e , g , i , k , o , p , r , s , u and v and a two-way ANOVA followed by a Bonferroni post hoc test in q . The data are presented as the mean ± s.e.m. The P values are presented on the graph.

Article Snippet: The primary antibodies used were: anti-Iba1 (1:100, ab283346, Abcam), anti-Iba1 (1:200, A19776, ABclonal), anti-NeuN (1:500, ab279296, Abcam), anti-NeuN (1:400, ab104224, Abcam), anti-MAP2 (1:200, 8707, Cell Signaling Technology), anti-AT8 (phospho-Tau (S202/T205), 1:600, GB113883 , Servicebio), anti-CD44 (1:500, GB112054 , Servicebio), anti-Hv1(1:100, AHC-001, Alomone Labs), anti-CD11c (1:100, ab254183, Abcam), anti-mouse I-A/I-E (1:100, 107601, BioLegend), anti-nicotinamide adenine dinucleotide phosphate oxidase 2 (NOX2) (1:200, GB11391-100, Servicebio), anti-GFAP (1:500, BD12096, Servicebio), anti-CD4 (1:100, 100505, BioLegend) and anti-CD8 (1:200, GB114196 , Servicebio) antibodies.

Techniques: Staining, Fluorescence

Journal: Experimental & Molecular Medicine

Article Title: Hv1 inhibition rescues AD pathology by restoring microglial mitochondrial function and enhancing mitochondrial transfer

doi: 10.1038/s12276-025-01593-z

Figure Lengend Snippet: a A GSEA analysis showing significantly downregulated mitochondria-associated pathways in AAV-TAU compared with AAV-CTRL. b GSEA plots depicting ‘electron transport activity’ and ‘respirasome’ pathways were enriched in AAV-CTRL compared with AAV-TAU. c A GSEA showing significantly upregulated mitochondria-associated pathways in AAV-TAU + YHV98-4 compared with AAV-TAU. d GSEA plots depicting enrichment of ‘electron transport activity’ and ‘ATP synthesis coupled electron transport’ in AAV-TAU + YHV98-4 compared with AAV-TAU. e Representative transmission electron microscopy images of PHF before and after sonication. f Representative images of MitoTracker (red) colabeled with Hv1 (green) and DAPI (blue) in BV2 cells. g , h A quantification of Hv1 intensity ( n = 48–50 cells from three independent experiments) ( g ) and MitoTracker fluorescence intensity ( h ) ( n = 12–14 fields of view from three independent experiments) in PHF-treated cells, with or without YHV98-4 treatment. i , j Measurement of CI activity ( i ) and CV activity ( j ) in cells treated with 1 μg/ml PHF, with or without 20 μM YHV98-4 ( n = 3 independent experiments). k A schematic diagram illustrating how Hv1 triggers ETC damage. CI, complex I; CV, complex V. The data were calculated using a one-way ANOVA followed by a Tukey’s post hoc analysis and presented as the mean ± s.e.m. The P values are presented on the graph.

Article Snippet: The primary antibodies used were: anti-Iba1 (1:100, ab283346, Abcam), anti-Iba1 (1:200, A19776, ABclonal), anti-NeuN (1:500, ab279296, Abcam), anti-NeuN (1:400, ab104224, Abcam), anti-MAP2 (1:200, 8707, Cell Signaling Technology), anti-AT8 (phospho-Tau (S202/T205), 1:600, GB113883 , Servicebio), anti-CD44 (1:500, GB112054 , Servicebio), anti-Hv1(1:100, AHC-001, Alomone Labs), anti-CD11c (1:100, ab254183, Abcam), anti-mouse I-A/I-E (1:100, 107601, BioLegend), anti-nicotinamide adenine dinucleotide phosphate oxidase 2 (NOX2) (1:200, GB11391-100, Servicebio), anti-GFAP (1:500, BD12096, Servicebio), anti-CD4 (1:100, 100505, BioLegend) and anti-CD8 (1:200, GB114196 , Servicebio) antibodies.

Techniques: Activity Assay, Transmission Assay, Electron Microscopy, Sonication, Fluorescence