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goat anti mouse cd45  (Santa Cruz Biotechnology)


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    Santa Cruz Biotechnology goat anti mouse cd45
    (A-B) Representative multiplex immunofluorescence images of an ovarian tumor from young or aged hosts. Tumor cells are identified as CK8⁺(yellow), immune cells as CD45⁺ (white), endothelial cells as CD31⁺(red), and nuclei (blue). Magnified panels highlight representative regions of interest (ROIs) selected from CD45⁺depleted or CD45⁺enriched areas for Spatial GeoMx analysis (n=3). (C) Intra-tumoral Gene Set Enrichment Analysis (GSEA) comparing CD45⁺depleted or CD45⁺-enriched regions from (i) young and (ii) aged hosts tumors using normalized enrichment scores (NES) against MSigDB Hallmark gene sets (FDR<0.05). (D) Heatmaps depicting unique inflammatory genes contributing to the ‘Inflammatory Hallmark’ enrichment in ovarian tumors from (i) young (ii) aged hosts. ( E) <t>CD45</t> + cells deconvolution using CIBERSORTX from young and aged host tumors. (F) (i-iv) GSEA of CD45⁺-enriched regions compared between young and aged hosts using Hallmark gene sets (FDR<0.05). (G) (i-iii) The top three upregulated genes in aged host ovarian tumors are associated with higher B cells and prolonged survival of high-grade ovarian cancer patients, and (iv-vi) The top three upregulated genes in aged host ovarian tumors are associated with higher M2 macrophages and lower survival of high-grade serous ovarian cancer patients.
    Goat Anti Mouse Cd45, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 876 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/goat+anti+cd45/CD45/bio_rxiv__64898__2025__12__23__695206-183-5-36
    Average 96 stars, based on 876 article reviews
    goat anti mouse cd45 - by Bioz Stars, 2026-10
    96/100 stars

    Images

    1) Product Images from "Aging increases ovarian cancer growth, metastasis, and immunosuppression that can be alleviated by inhibiting hedgehog signaling"

    Article Title: Aging increases ovarian cancer growth, metastasis, and immunosuppression that can be alleviated by inhibiting hedgehog signaling

    Journal: bioRxiv

    doi: 10.64898/2025.12.23.695206

    (A-B) Representative multiplex immunofluorescence images of an ovarian tumor from young or aged hosts. Tumor cells are identified as CK8⁺(yellow), immune cells as CD45⁺ (white), endothelial cells as CD31⁺(red), and nuclei (blue). Magnified panels highlight representative regions of interest (ROIs) selected from CD45⁺depleted or CD45⁺enriched areas for Spatial GeoMx analysis (n=3). (C) Intra-tumoral Gene Set Enrichment Analysis (GSEA) comparing CD45⁺depleted or CD45⁺-enriched regions from (i) young and (ii) aged hosts tumors using normalized enrichment scores (NES) against MSigDB Hallmark gene sets (FDR<0.05). (D) Heatmaps depicting unique inflammatory genes contributing to the ‘Inflammatory Hallmark’ enrichment in ovarian tumors from (i) young (ii) aged hosts. ( E) CD45 + cells deconvolution using CIBERSORTX from young and aged host tumors. (F) (i-iv) GSEA of CD45⁺-enriched regions compared between young and aged hosts using Hallmark gene sets (FDR<0.05). (G) (i-iii) The top three upregulated genes in aged host ovarian tumors are associated with higher B cells and prolonged survival of high-grade ovarian cancer patients, and (iv-vi) The top three upregulated genes in aged host ovarian tumors are associated with higher M2 macrophages and lower survival of high-grade serous ovarian cancer patients.
    Figure Legend Snippet: (A-B) Representative multiplex immunofluorescence images of an ovarian tumor from young or aged hosts. Tumor cells are identified as CK8⁺(yellow), immune cells as CD45⁺ (white), endothelial cells as CD31⁺(red), and nuclei (blue). Magnified panels highlight representative regions of interest (ROIs) selected from CD45⁺depleted or CD45⁺enriched areas for Spatial GeoMx analysis (n=3). (C) Intra-tumoral Gene Set Enrichment Analysis (GSEA) comparing CD45⁺depleted or CD45⁺-enriched regions from (i) young and (ii) aged hosts tumors using normalized enrichment scores (NES) against MSigDB Hallmark gene sets (FDR<0.05). (D) Heatmaps depicting unique inflammatory genes contributing to the ‘Inflammatory Hallmark’ enrichment in ovarian tumors from (i) young (ii) aged hosts. ( E) CD45 + cells deconvolution using CIBERSORTX from young and aged host tumors. (F) (i-iv) GSEA of CD45⁺-enriched regions compared between young and aged hosts using Hallmark gene sets (FDR<0.05). (G) (i-iii) The top three upregulated genes in aged host ovarian tumors are associated with higher B cells and prolonged survival of high-grade ovarian cancer patients, and (iv-vi) The top three upregulated genes in aged host ovarian tumors are associated with higher M2 macrophages and lower survival of high-grade serous ovarian cancer patients.

    Techniques Used: Multiplex Assay, Immunofluorescence

    (A-B) Representative immunofluorescence images from ovarian tumors (A) (i) ID8 Trp53 −/− (B) (i) PPNM cells in ovarian tumors showing CD45 + (green), Arg1 + (red), and DAPI (blue). Adjacent graphs ( A) (ii) and (B) (ii) showing quantification of percentage CD45 + Arg1 + cells in ovarian tumors from both models. (C-D) Representative immunofluorescence images from (C) (i) ID8 Trp53 −/− (D) (i) PPNM cells in ovarian tumors showing CD68 + (green), Arg1 + (red), and DAPI (blue) cells. Quantification of percentage CD68 + Arg1 + cells (C) (ii) and (D) (ii) showing ovarian tumors from both models. (E-F) Immunohistochemistry of ovarian tumors showing CD206 + cells (E)( i) ID8 Trp53 −/− (F) (i) PPNM cells in ovarian tumors. Quantification of percentage CD206 + cells. (E) (ii) and (F) (ii) in both models. (G-H) Representative immunofluorescence images of Foxp3 + (red) and DAPI (blue) immunostaining( G) (i) ID8 Trp53 −/− (H) (i) PPNM in ovarian tumor. Graphs (G) (ii) and (H) (ii) showing quantification of percentage Foxp3 + cells in both models. (I-J) Representative immunofluorescence images from (I) (i) ID8 Trp53 −/− (J) (i) PPNM cells in omental tumors showing CD45 + (green), Arg1 + (red), and DAPI(blue). Adjacent graphs (I) (ii) and (J) (ii) showing quantification of percentage CD45 + Arg1 + cells in omental tumors from both models. (K-L) Representative immunofluorescence images from (K) (i) ID8 Trp53 −/− (L) (i) PPNM cells in omental tumors showing CD68 + (green), Arg1 + (red), and DAPI (blue) cells. (K) (ii) and (L) (ii) Quantification of percentage CD68 + Arg1 + cells showing omental tumors from both models in (p=0.169 and p<0.05, respectively). (M-N) Immunohistochemistry images of omental tumors showing CD206 + cells (M) (i) ID8 Trp53 −/− (N) (i) PPNM cells in omental tumors. (M) (ii) and (N) (ii) Quantification of percentage CD206 + cells in both models. (O-P) Representative immunofluorescence images of Foxp3 + (red) and DAPI (blue) immunostaining ( O) (i) ID8 Trp53 −/− (P) (i) PPNM in omental tumor. Adjacent graphs (O) (ii) and (P) (ii) showing quantification of percentage Foxp3 + cells in both models. Color codes and symbols indicate the number of images analyzed per mouse in each group. All data are meant SEM (n=3), *p<0.05, **p<0.01, *** p<0.001, ****p<0.0001, unpaired t test.
    Figure Legend Snippet: (A-B) Representative immunofluorescence images from ovarian tumors (A) (i) ID8 Trp53 −/− (B) (i) PPNM cells in ovarian tumors showing CD45 + (green), Arg1 + (red), and DAPI (blue). Adjacent graphs ( A) (ii) and (B) (ii) showing quantification of percentage CD45 + Arg1 + cells in ovarian tumors from both models. (C-D) Representative immunofluorescence images from (C) (i) ID8 Trp53 −/− (D) (i) PPNM cells in ovarian tumors showing CD68 + (green), Arg1 + (red), and DAPI (blue) cells. Quantification of percentage CD68 + Arg1 + cells (C) (ii) and (D) (ii) showing ovarian tumors from both models. (E-F) Immunohistochemistry of ovarian tumors showing CD206 + cells (E)( i) ID8 Trp53 −/− (F) (i) PPNM cells in ovarian tumors. Quantification of percentage CD206 + cells. (E) (ii) and (F) (ii) in both models. (G-H) Representative immunofluorescence images of Foxp3 + (red) and DAPI (blue) immunostaining( G) (i) ID8 Trp53 −/− (H) (i) PPNM in ovarian tumor. Graphs (G) (ii) and (H) (ii) showing quantification of percentage Foxp3 + cells in both models. (I-J) Representative immunofluorescence images from (I) (i) ID8 Trp53 −/− (J) (i) PPNM cells in omental tumors showing CD45 + (green), Arg1 + (red), and DAPI(blue). Adjacent graphs (I) (ii) and (J) (ii) showing quantification of percentage CD45 + Arg1 + cells in omental tumors from both models. (K-L) Representative immunofluorescence images from (K) (i) ID8 Trp53 −/− (L) (i) PPNM cells in omental tumors showing CD68 + (green), Arg1 + (red), and DAPI (blue) cells. (K) (ii) and (L) (ii) Quantification of percentage CD68 + Arg1 + cells showing omental tumors from both models in (p=0.169 and p<0.05, respectively). (M-N) Immunohistochemistry images of omental tumors showing CD206 + cells (M) (i) ID8 Trp53 −/− (N) (i) PPNM cells in omental tumors. (M) (ii) and (N) (ii) Quantification of percentage CD206 + cells in both models. (O-P) Representative immunofluorescence images of Foxp3 + (red) and DAPI (blue) immunostaining ( O) (i) ID8 Trp53 −/− (P) (i) PPNM in omental tumor. Adjacent graphs (O) (ii) and (P) (ii) showing quantification of percentage Foxp3 + cells in both models. Color codes and symbols indicate the number of images analyzed per mouse in each group. All data are meant SEM (n=3), *p<0.05, **p<0.01, *** p<0.001, ****p<0.0001, unpaired t test.

    Techniques Used: Immunofluorescence, Immunohistochemistry, Immunostaining

    (A-B) Representative immunofluorescence images of CD45 + (green), Arg1 + (red), and DAPI (blue) in (A) (i) ovarian (B) (i) omental tumors from vismodegib and vehicle treated mice. Adjacent graphs showing percentage of CD45 + Arg1 + cells in (A) (ii) ovarian tumors (p=0.062) and (B) (ii) in omentum (p=0.372). (C-D) Representative immunofluorescence images of CD68 + (green), Arg1 + (red), and DAPI (Blue) in (C) (i) ovarian or (D) (i) omental tumors from vismodegib and vehicle treated mice. Adjacent graphs showing percentage of CD68 + Arg1 + cells in (C) (ii) ovaries (p=0.502), (D) (ii) in omentum (p=0.317). (E-F) Immunohistochemistry of CD206 + cells in (E) (i) ovarian ( F )(i) omental tumors. Adjacent graph showing percentage CD206 + cells (E) (ii) in ovaries and (F) (ii) in omentum (p=0.624). (G-H) Representative immunofluorescence images of Foxp3 + (red) and DAPI (Blue) cells ( G) (i) ovarian (H) (i) omental tumors. Adjacent graph showing percentage Foxp3 + cells in ( G) (ii) ovarian and (H) (ii) omental tumors. Color codes and symbols indicate the number of images analyzed per mouse in each group. All data are presented as mean±SEM (n=3), *p<0.05, **p<0.01, ***p<0.001, ***p<0.0001, unpaired t test.
    Figure Legend Snippet: (A-B) Representative immunofluorescence images of CD45 + (green), Arg1 + (red), and DAPI (blue) in (A) (i) ovarian (B) (i) omental tumors from vismodegib and vehicle treated mice. Adjacent graphs showing percentage of CD45 + Arg1 + cells in (A) (ii) ovarian tumors (p=0.062) and (B) (ii) in omentum (p=0.372). (C-D) Representative immunofluorescence images of CD68 + (green), Arg1 + (red), and DAPI (Blue) in (C) (i) ovarian or (D) (i) omental tumors from vismodegib and vehicle treated mice. Adjacent graphs showing percentage of CD68 + Arg1 + cells in (C) (ii) ovaries (p=0.502), (D) (ii) in omentum (p=0.317). (E-F) Immunohistochemistry of CD206 + cells in (E) (i) ovarian ( F )(i) omental tumors. Adjacent graph showing percentage CD206 + cells (E) (ii) in ovaries and (F) (ii) in omentum (p=0.624). (G-H) Representative immunofluorescence images of Foxp3 + (red) and DAPI (Blue) cells ( G) (i) ovarian (H) (i) omental tumors. Adjacent graph showing percentage Foxp3 + cells in ( G) (ii) ovarian and (H) (ii) omental tumors. Color codes and symbols indicate the number of images analyzed per mouse in each group. All data are presented as mean±SEM (n=3), *p<0.05, **p<0.01, ***p<0.001, ***p<0.0001, unpaired t test.

    Techniques Used: Immunofluorescence, Immunohistochemistry

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    Staining:

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    Incubation:

    Article Title: Adipose-Derived Stem Cell Treatment Induces Early-Term Hes1 Upregulation in a Sox9- and Notch1-Independent Manner in a Rat Model of Bile Duct Ligation.
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    Immunohistochemical staining:

    Article Title: Adipose-Derived Stem Cell Treatment Induces Early-Term Hes1 Upregulation in a Sox9- and Notch1-Independent Manner in a Rat Model of Bile Duct Ligation.
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    Immunohistochemistry:

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    Cell-Signaling:

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    Immunofluorescence:

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    Expressing:

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    Article Snippet: .. To selectively evaluate the expression of key cellular markers, immunofluorescence and flow cytometry were performed according to standard protocols on a confocal fluorescence microscope (Leica TCS SP8, Germany) or a flow cytometer (BD LSRFortessa X-20 cell analyzer, USA), respectively, using the following immunofluorescence antibodies: CD29, CD90, CD105, CD45 (mesenchymal stromal cell marker, Abcam, ab93758), STRO-1 (Abcam, AB214086 ), CD34 (Abcam, ab81289), and CD73 (Abcam, ab317462); and flow cytometer antibodies: CD29 (BD, 561794), CD90 (BD, 561972), CD105 (BD, 560839), STRO-1 (Santa Cruz, sc-47733), CD34 (BD, 561440), CD45 (BD, 560915), and CD73 (BD, 561014) in concentrations recommended by the manufacturer. ..

    Flow Cytometry:

    Article Title: A Native Bioactive Interface Functionalized with Osteoprogenitor Stem Cell-Derived Migrasomes for Enhanced Bone Regeneration
    Article Snippet: .. To selectively evaluate the expression of key cellular markers, immunofluorescence and flow cytometry were performed according to standard protocols on a confocal fluorescence microscope (Leica TCS SP8, Germany) or a flow cytometer (BD LSRFortessa X-20 cell analyzer, USA), respectively, using the following immunofluorescence antibodies: CD29, CD90, CD105, CD45 (mesenchymal stromal cell marker, Abcam, ab93758), STRO-1 (Abcam, AB214086 ), CD34 (Abcam, ab81289), and CD73 (Abcam, ab317462); and flow cytometer antibodies: CD29 (BD, 561794), CD90 (BD, 561972), CD105 (BD, 560839), STRO-1 (Santa Cruz, sc-47733), CD34 (BD, 561440), CD45 (BD, 560915), and CD73 (BD, 561014) in concentrations recommended by the manufacturer. ..

    Confocal:

    Article Title: A Native Bioactive Interface Functionalized with Osteoprogenitor Stem Cell-Derived Migrasomes for Enhanced Bone Regeneration
    Article Snippet: .. To selectively evaluate the expression of key cellular markers, immunofluorescence and flow cytometry were performed according to standard protocols on a confocal fluorescence microscope (Leica TCS SP8, Germany) or a flow cytometer (BD LSRFortessa X-20 cell analyzer, USA), respectively, using the following immunofluorescence antibodies: CD29, CD90, CD105, CD45 (mesenchymal stromal cell marker, Abcam, ab93758), STRO-1 (Abcam, AB214086 ), CD34 (Abcam, ab81289), and CD73 (Abcam, ab317462); and flow cytometer antibodies: CD29 (BD, 561794), CD90 (BD, 561972), CD105 (BD, 560839), STRO-1 (Santa Cruz, sc-47733), CD34 (BD, 561440), CD45 (BD, 560915), and CD73 (BD, 561014) in concentrations recommended by the manufacturer. ..

    Fluorescence:

    Article Title: A Native Bioactive Interface Functionalized with Osteoprogenitor Stem Cell-Derived Migrasomes for Enhanced Bone Regeneration
    Article Snippet: .. To selectively evaluate the expression of key cellular markers, immunofluorescence and flow cytometry were performed according to standard protocols on a confocal fluorescence microscope (Leica TCS SP8, Germany) or a flow cytometer (BD LSRFortessa X-20 cell analyzer, USA), respectively, using the following immunofluorescence antibodies: CD29, CD90, CD105, CD45 (mesenchymal stromal cell marker, Abcam, ab93758), STRO-1 (Abcam, AB214086 ), CD34 (Abcam, ab81289), and CD73 (Abcam, ab317462); and flow cytometer antibodies: CD29 (BD, 561794), CD90 (BD, 561972), CD105 (BD, 560839), STRO-1 (Santa Cruz, sc-47733), CD34 (BD, 561440), CD45 (BD, 560915), and CD73 (BD, 561014) in concentrations recommended by the manufacturer. ..

    Microscopy:

    Article Title: A Native Bioactive Interface Functionalized with Osteoprogenitor Stem Cell-Derived Migrasomes for Enhanced Bone Regeneration
    Article Snippet: .. To selectively evaluate the expression of key cellular markers, immunofluorescence and flow cytometry were performed according to standard protocols on a confocal fluorescence microscope (Leica TCS SP8, Germany) or a flow cytometer (BD LSRFortessa X-20 cell analyzer, USA), respectively, using the following immunofluorescence antibodies: CD29, CD90, CD105, CD45 (mesenchymal stromal cell marker, Abcam, ab93758), STRO-1 (Abcam, AB214086 ), CD34 (Abcam, ab81289), and CD73 (Abcam, ab317462); and flow cytometer antibodies: CD29 (BD, 561794), CD90 (BD, 561972), CD105 (BD, 560839), STRO-1 (Santa Cruz, sc-47733), CD34 (BD, 561440), CD45 (BD, 560915), and CD73 (BD, 561014) in concentrations recommended by the manufacturer. ..

    Marker:

    Article Title: A Native Bioactive Interface Functionalized with Osteoprogenitor Stem Cell-Derived Migrasomes for Enhanced Bone Regeneration
    Article Snippet: .. To selectively evaluate the expression of key cellular markers, immunofluorescence and flow cytometry were performed according to standard protocols on a confocal fluorescence microscope (Leica TCS SP8, Germany) or a flow cytometer (BD LSRFortessa X-20 cell analyzer, USA), respectively, using the following immunofluorescence antibodies: CD29, CD90, CD105, CD45 (mesenchymal stromal cell marker, Abcam, ab93758), STRO-1 (Abcam, AB214086 ), CD34 (Abcam, ab81289), and CD73 (Abcam, ab317462); and flow cytometer antibodies: CD29 (BD, 561794), CD90 (BD, 561972), CD105 (BD, 560839), STRO-1 (Santa Cruz, sc-47733), CD34 (BD, 561440), CD45 (BD, 560915), and CD73 (BD, 561014) in concentrations recommended by the manufacturer. ..



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    96
    Santa Cruz Biotechnology goat anti mouse cd45
    (A-B) Representative multiplex immunofluorescence images of an ovarian tumor from young or aged hosts. Tumor cells are identified as CK8⁺(yellow), immune cells as CD45⁺ (white), endothelial cells as CD31⁺(red), and nuclei (blue). Magnified panels highlight representative regions of interest (ROIs) selected from CD45⁺depleted or CD45⁺enriched areas for Spatial GeoMx analysis (n=3). (C) Intra-tumoral Gene Set Enrichment Analysis (GSEA) comparing CD45⁺depleted or CD45⁺-enriched regions from (i) young and (ii) aged hosts tumors using normalized enrichment scores (NES) against MSigDB Hallmark gene sets (FDR<0.05). (D) Heatmaps depicting unique inflammatory genes contributing to the ‘Inflammatory Hallmark’ enrichment in ovarian tumors from (i) young (ii) aged hosts. ( E) <t>CD45</t> + cells deconvolution using CIBERSORTX from young and aged host tumors. (F) (i-iv) GSEA of CD45⁺-enriched regions compared between young and aged hosts using Hallmark gene sets (FDR<0.05). (G) (i-iii) The top three upregulated genes in aged host ovarian tumors are associated with higher B cells and prolonged survival of high-grade ovarian cancer patients, and (iv-vi) The top three upregulated genes in aged host ovarian tumors are associated with higher M2 macrophages and lower survival of high-grade serous ovarian cancer patients.
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    ( A ) Cdh5CreER;Tgfbr1 CKO/- retinas showing CNV (white arrows in central panels) and vascular invasion of the outer nuclear layer (white arrows in lower panels), both with associated <t>CD45+</t> immune cells. ( B ) Cdh5CreER;Tgfbr2 CKO/- retinas showing CNV (white arrows in central panels) and an anastomosis between retinal and choroidal vasculatures (white arrows in lower panels), with intraretinal EC marker CLDN5, choroidal EC marker PLVAP, and pan-vascular marker COL4 (collagen4). ( C ) Upper right panels, in the Cdh5CreER;Tgfbr1 CKO/- retina, CNV (white arrows) is derived from choroidal vasculature, marked by PLVAP. Lower right panels, CNV (white arrows) is present in the subretinal space, i.e., on the retinal side of the RPE, which is marked by RPE65. Abbreviations: Ch., choroid; ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer; RPE, retinal pigment epithelium. The ages of the mice are indicated in postnatal days (P) for this and all other figures. Scale bars, 100 µm.
    Goat Polyclonal Anti Mouse Cd45, supplied by R&D Systems, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ( A ) Cdh5CreER;Tgfbr1 CKO/- retinas showing CNV (white arrows in central panels) and vascular invasion of the outer nuclear layer (white arrows in lower panels), both with associated <t>CD45+</t> immune cells. ( B ) Cdh5CreER;Tgfbr2 CKO/- retinas showing CNV (white arrows in central panels) and an anastomosis between retinal and choroidal vasculatures (white arrows in lower panels), with intraretinal EC marker CLDN5, choroidal EC marker PLVAP, and pan-vascular marker COL4 (collagen4). ( C ) Upper right panels, in the Cdh5CreER;Tgfbr1 CKO/- retina, CNV (white arrows) is derived from choroidal vasculature, marked by PLVAP. Lower right panels, CNV (white arrows) is present in the subretinal space, i.e., on the retinal side of the RPE, which is marked by RPE65. Abbreviations: Ch., choroid; ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer; RPE, retinal pigment epithelium. The ages of the mice are indicated in postnatal days (P) for this and all other figures. Scale bars, 100 µm.
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    ( A ) Cdh5CreER;Tgfbr1 CKO/- retinas showing CNV (white arrows in central panels) and vascular invasion of the outer nuclear layer (white arrows in lower panels), both with associated <t>CD45+</t> immune cells. ( B ) Cdh5CreER;Tgfbr2 CKO/- retinas showing CNV (white arrows in central panels) and an anastomosis between retinal and choroidal vasculatures (white arrows in lower panels), with intraretinal EC marker CLDN5, choroidal EC marker PLVAP, and pan-vascular marker COL4 (collagen4). ( C ) Upper right panels, in the Cdh5CreER;Tgfbr1 CKO/- retina, CNV (white arrows) is derived from choroidal vasculature, marked by PLVAP. Lower right panels, CNV (white arrows) is present in the subretinal space, i.e., on the retinal side of the RPE, which is marked by RPE65. Abbreviations: Ch., choroid; ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer; RPE, retinal pigment epithelium. The ages of the mice are indicated in postnatal days (P) for this and all other figures. Scale bars, 100 µm.
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    ( A ) Cdh5CreER;Tgfbr1 CKO/- retinas showing CNV (white arrows in central panels) and vascular invasion of the outer nuclear layer (white arrows in lower panels), both with associated <t>CD45+</t> immune cells. ( B ) Cdh5CreER;Tgfbr2 CKO/- retinas showing CNV (white arrows in central panels) and an anastomosis between retinal and choroidal vasculatures (white arrows in lower panels), with intraretinal EC marker CLDN5, choroidal EC marker PLVAP, and pan-vascular marker COL4 (collagen4). ( C ) Upper right panels, in the Cdh5CreER;Tgfbr1 CKO/- retina, CNV (white arrows) is derived from choroidal vasculature, marked by PLVAP. Lower right panels, CNV (white arrows) is present in the subretinal space, i.e., on the retinal side of the RPE, which is marked by RPE65. Abbreviations: Ch., choroid; ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer; RPE, retinal pigment epithelium. The ages of the mice are indicated in postnatal days (P) for this and all other figures. Scale bars, 100 µm.
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    Pharmacological inhibition of IL12β attenuated TAC-induced LV inflammation. (A, B) Representative images and quantified data of LV <t>CD45</t> + leukocytes performed by immuno-histological staining. (C) Flow cytometry plot for the identification of CD45 + leukocytes in the LV. (D) Quantified data of the number of CD45 + leukocytes per LV. (E) The percentage of immune cell subsets within CD45 + leukocytes. (F) Quantified data of number of different immune cell subsets per LV. *p<0.05; # p<0.05 IgG-treated TAC mice compared with the control; † p<0.05 anti-IL12β-treated TAC mice compared with IgG-treated TAC mice; Neutro, Neutrophils; Mφ, Macrophage; Mono, Monocytes; DCs, Dendritic Cells; NK, Natural Killer Cells; n = 3–5 mice per group.
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    Pharmacological inhibition of IL12β attenuated TAC-induced LV inflammation. (A, B) Representative images and quantified data of LV <t>CD45</t> + leukocytes performed by immuno-histological staining. (C) Flow cytometry plot for the identification of CD45 + leukocytes in the LV. (D) Quantified data of the number of CD45 + leukocytes per LV. (E) The percentage of immune cell subsets within CD45 + leukocytes. (F) Quantified data of number of different immune cell subsets per LV. *p<0.05; # p<0.05 IgG-treated TAC mice compared with the control; † p<0.05 anti-IL12β-treated TAC mice compared with IgG-treated TAC mice; Neutro, Neutrophils; Mφ, Macrophage; Mono, Monocytes; DCs, Dendritic Cells; NK, Natural Killer Cells; n = 3–5 mice per group.
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    Image Search Results


    (A-B) Representative multiplex immunofluorescence images of an ovarian tumor from young or aged hosts. Tumor cells are identified as CK8⁺(yellow), immune cells as CD45⁺ (white), endothelial cells as CD31⁺(red), and nuclei (blue). Magnified panels highlight representative regions of interest (ROIs) selected from CD45⁺depleted or CD45⁺enriched areas for Spatial GeoMx analysis (n=3). (C) Intra-tumoral Gene Set Enrichment Analysis (GSEA) comparing CD45⁺depleted or CD45⁺-enriched regions from (i) young and (ii) aged hosts tumors using normalized enrichment scores (NES) against MSigDB Hallmark gene sets (FDR<0.05). (D) Heatmaps depicting unique inflammatory genes contributing to the ‘Inflammatory Hallmark’ enrichment in ovarian tumors from (i) young (ii) aged hosts. ( E) CD45 + cells deconvolution using CIBERSORTX from young and aged host tumors. (F) (i-iv) GSEA of CD45⁺-enriched regions compared between young and aged hosts using Hallmark gene sets (FDR<0.05). (G) (i-iii) The top three upregulated genes in aged host ovarian tumors are associated with higher B cells and prolonged survival of high-grade ovarian cancer patients, and (iv-vi) The top three upregulated genes in aged host ovarian tumors are associated with higher M2 macrophages and lower survival of high-grade serous ovarian cancer patients.

    Journal: bioRxiv

    Article Title: Aging increases ovarian cancer growth, metastasis, and immunosuppression that can be alleviated by inhibiting hedgehog signaling

    doi: 10.64898/2025.12.23.695206

    Figure Lengend Snippet: (A-B) Representative multiplex immunofluorescence images of an ovarian tumor from young or aged hosts. Tumor cells are identified as CK8⁺(yellow), immune cells as CD45⁺ (white), endothelial cells as CD31⁺(red), and nuclei (blue). Magnified panels highlight representative regions of interest (ROIs) selected from CD45⁺depleted or CD45⁺enriched areas for Spatial GeoMx analysis (n=3). (C) Intra-tumoral Gene Set Enrichment Analysis (GSEA) comparing CD45⁺depleted or CD45⁺-enriched regions from (i) young and (ii) aged hosts tumors using normalized enrichment scores (NES) against MSigDB Hallmark gene sets (FDR<0.05). (D) Heatmaps depicting unique inflammatory genes contributing to the ‘Inflammatory Hallmark’ enrichment in ovarian tumors from (i) young (ii) aged hosts. ( E) CD45 + cells deconvolution using CIBERSORTX from young and aged host tumors. (F) (i-iv) GSEA of CD45⁺-enriched regions compared between young and aged hosts using Hallmark gene sets (FDR<0.05). (G) (i-iii) The top three upregulated genes in aged host ovarian tumors are associated with higher B cells and prolonged survival of high-grade ovarian cancer patients, and (iv-vi) The top three upregulated genes in aged host ovarian tumors are associated with higher M2 macrophages and lower survival of high-grade serous ovarian cancer patients.

    Article Snippet: They were stained with labeled Goat anti-mouse CD45 (Cat no. AF114 1:100, Novus biologicals) primary antibody using FITC Conjugation Kit (Cat no. ab188285, Abcam) and Labeled Rat anti-mouse CK8 primary antibody (Cat no. Sc 8010, 1:100, Santa Cruz Biotechnology) using APC Conjugation Kit (Cat no. ab201807, Abcam) for immune and tumor cells detection respectively.

    Techniques: Multiplex Assay, Immunofluorescence

    (A-B) Representative immunofluorescence images from ovarian tumors (A) (i) ID8 Trp53 −/− (B) (i) PPNM cells in ovarian tumors showing CD45 + (green), Arg1 + (red), and DAPI (blue). Adjacent graphs ( A) (ii) and (B) (ii) showing quantification of percentage CD45 + Arg1 + cells in ovarian tumors from both models. (C-D) Representative immunofluorescence images from (C) (i) ID8 Trp53 −/− (D) (i) PPNM cells in ovarian tumors showing CD68 + (green), Arg1 + (red), and DAPI (blue) cells. Quantification of percentage CD68 + Arg1 + cells (C) (ii) and (D) (ii) showing ovarian tumors from both models. (E-F) Immunohistochemistry of ovarian tumors showing CD206 + cells (E)( i) ID8 Trp53 −/− (F) (i) PPNM cells in ovarian tumors. Quantification of percentage CD206 + cells. (E) (ii) and (F) (ii) in both models. (G-H) Representative immunofluorescence images of Foxp3 + (red) and DAPI (blue) immunostaining( G) (i) ID8 Trp53 −/− (H) (i) PPNM in ovarian tumor. Graphs (G) (ii) and (H) (ii) showing quantification of percentage Foxp3 + cells in both models. (I-J) Representative immunofluorescence images from (I) (i) ID8 Trp53 −/− (J) (i) PPNM cells in omental tumors showing CD45 + (green), Arg1 + (red), and DAPI(blue). Adjacent graphs (I) (ii) and (J) (ii) showing quantification of percentage CD45 + Arg1 + cells in omental tumors from both models. (K-L) Representative immunofluorescence images from (K) (i) ID8 Trp53 −/− (L) (i) PPNM cells in omental tumors showing CD68 + (green), Arg1 + (red), and DAPI (blue) cells. (K) (ii) and (L) (ii) Quantification of percentage CD68 + Arg1 + cells showing omental tumors from both models in (p=0.169 and p<0.05, respectively). (M-N) Immunohistochemistry images of omental tumors showing CD206 + cells (M) (i) ID8 Trp53 −/− (N) (i) PPNM cells in omental tumors. (M) (ii) and (N) (ii) Quantification of percentage CD206 + cells in both models. (O-P) Representative immunofluorescence images of Foxp3 + (red) and DAPI (blue) immunostaining ( O) (i) ID8 Trp53 −/− (P) (i) PPNM in omental tumor. Adjacent graphs (O) (ii) and (P) (ii) showing quantification of percentage Foxp3 + cells in both models. Color codes and symbols indicate the number of images analyzed per mouse in each group. All data are meant SEM (n=3), *p<0.05, **p<0.01, *** p<0.001, ****p<0.0001, unpaired t test.

    Journal: bioRxiv

    Article Title: Aging increases ovarian cancer growth, metastasis, and immunosuppression that can be alleviated by inhibiting hedgehog signaling

    doi: 10.64898/2025.12.23.695206

    Figure Lengend Snippet: (A-B) Representative immunofluorescence images from ovarian tumors (A) (i) ID8 Trp53 −/− (B) (i) PPNM cells in ovarian tumors showing CD45 + (green), Arg1 + (red), and DAPI (blue). Adjacent graphs ( A) (ii) and (B) (ii) showing quantification of percentage CD45 + Arg1 + cells in ovarian tumors from both models. (C-D) Representative immunofluorescence images from (C) (i) ID8 Trp53 −/− (D) (i) PPNM cells in ovarian tumors showing CD68 + (green), Arg1 + (red), and DAPI (blue) cells. Quantification of percentage CD68 + Arg1 + cells (C) (ii) and (D) (ii) showing ovarian tumors from both models. (E-F) Immunohistochemistry of ovarian tumors showing CD206 + cells (E)( i) ID8 Trp53 −/− (F) (i) PPNM cells in ovarian tumors. Quantification of percentage CD206 + cells. (E) (ii) and (F) (ii) in both models. (G-H) Representative immunofluorescence images of Foxp3 + (red) and DAPI (blue) immunostaining( G) (i) ID8 Trp53 −/− (H) (i) PPNM in ovarian tumor. Graphs (G) (ii) and (H) (ii) showing quantification of percentage Foxp3 + cells in both models. (I-J) Representative immunofluorescence images from (I) (i) ID8 Trp53 −/− (J) (i) PPNM cells in omental tumors showing CD45 + (green), Arg1 + (red), and DAPI(blue). Adjacent graphs (I) (ii) and (J) (ii) showing quantification of percentage CD45 + Arg1 + cells in omental tumors from both models. (K-L) Representative immunofluorescence images from (K) (i) ID8 Trp53 −/− (L) (i) PPNM cells in omental tumors showing CD68 + (green), Arg1 + (red), and DAPI (blue) cells. (K) (ii) and (L) (ii) Quantification of percentage CD68 + Arg1 + cells showing omental tumors from both models in (p=0.169 and p<0.05, respectively). (M-N) Immunohistochemistry images of omental tumors showing CD206 + cells (M) (i) ID8 Trp53 −/− (N) (i) PPNM cells in omental tumors. (M) (ii) and (N) (ii) Quantification of percentage CD206 + cells in both models. (O-P) Representative immunofluorescence images of Foxp3 + (red) and DAPI (blue) immunostaining ( O) (i) ID8 Trp53 −/− (P) (i) PPNM in omental tumor. Adjacent graphs (O) (ii) and (P) (ii) showing quantification of percentage Foxp3 + cells in both models. Color codes and symbols indicate the number of images analyzed per mouse in each group. All data are meant SEM (n=3), *p<0.05, **p<0.01, *** p<0.001, ****p<0.0001, unpaired t test.

    Article Snippet: They were stained with labeled Goat anti-mouse CD45 (Cat no. AF114 1:100, Novus biologicals) primary antibody using FITC Conjugation Kit (Cat no. ab188285, Abcam) and Labeled Rat anti-mouse CK8 primary antibody (Cat no. Sc 8010, 1:100, Santa Cruz Biotechnology) using APC Conjugation Kit (Cat no. ab201807, Abcam) for immune and tumor cells detection respectively.

    Techniques: Immunofluorescence, Immunohistochemistry, Immunostaining

    (A-B) Representative immunofluorescence images of CD45 + (green), Arg1 + (red), and DAPI (blue) in (A) (i) ovarian (B) (i) omental tumors from vismodegib and vehicle treated mice. Adjacent graphs showing percentage of CD45 + Arg1 + cells in (A) (ii) ovarian tumors (p=0.062) and (B) (ii) in omentum (p=0.372). (C-D) Representative immunofluorescence images of CD68 + (green), Arg1 + (red), and DAPI (Blue) in (C) (i) ovarian or (D) (i) omental tumors from vismodegib and vehicle treated mice. Adjacent graphs showing percentage of CD68 + Arg1 + cells in (C) (ii) ovaries (p=0.502), (D) (ii) in omentum (p=0.317). (E-F) Immunohistochemistry of CD206 + cells in (E) (i) ovarian ( F )(i) omental tumors. Adjacent graph showing percentage CD206 + cells (E) (ii) in ovaries and (F) (ii) in omentum (p=0.624). (G-H) Representative immunofluorescence images of Foxp3 + (red) and DAPI (Blue) cells ( G) (i) ovarian (H) (i) omental tumors. Adjacent graph showing percentage Foxp3 + cells in ( G) (ii) ovarian and (H) (ii) omental tumors. Color codes and symbols indicate the number of images analyzed per mouse in each group. All data are presented as mean±SEM (n=3), *p<0.05, **p<0.01, ***p<0.001, ***p<0.0001, unpaired t test.

    Journal: bioRxiv

    Article Title: Aging increases ovarian cancer growth, metastasis, and immunosuppression that can be alleviated by inhibiting hedgehog signaling

    doi: 10.64898/2025.12.23.695206

    Figure Lengend Snippet: (A-B) Representative immunofluorescence images of CD45 + (green), Arg1 + (red), and DAPI (blue) in (A) (i) ovarian (B) (i) omental tumors from vismodegib and vehicle treated mice. Adjacent graphs showing percentage of CD45 + Arg1 + cells in (A) (ii) ovarian tumors (p=0.062) and (B) (ii) in omentum (p=0.372). (C-D) Representative immunofluorescence images of CD68 + (green), Arg1 + (red), and DAPI (Blue) in (C) (i) ovarian or (D) (i) omental tumors from vismodegib and vehicle treated mice. Adjacent graphs showing percentage of CD68 + Arg1 + cells in (C) (ii) ovaries (p=0.502), (D) (ii) in omentum (p=0.317). (E-F) Immunohistochemistry of CD206 + cells in (E) (i) ovarian ( F )(i) omental tumors. Adjacent graph showing percentage CD206 + cells (E) (ii) in ovaries and (F) (ii) in omentum (p=0.624). (G-H) Representative immunofluorescence images of Foxp3 + (red) and DAPI (Blue) cells ( G) (i) ovarian (H) (i) omental tumors. Adjacent graph showing percentage Foxp3 + cells in ( G) (ii) ovarian and (H) (ii) omental tumors. Color codes and symbols indicate the number of images analyzed per mouse in each group. All data are presented as mean±SEM (n=3), *p<0.05, **p<0.01, ***p<0.001, ***p<0.0001, unpaired t test.

    Article Snippet: They were stained with labeled Goat anti-mouse CD45 (Cat no. AF114 1:100, Novus biologicals) primary antibody using FITC Conjugation Kit (Cat no. ab188285, Abcam) and Labeled Rat anti-mouse CK8 primary antibody (Cat no. Sc 8010, 1:100, Santa Cruz Biotechnology) using APC Conjugation Kit (Cat no. ab201807, Abcam) for immune and tumor cells detection respectively.

    Techniques: Immunofluorescence, Immunohistochemistry

    ( A ) Cdh5CreER;Tgfbr1 CKO/- retinas showing CNV (white arrows in central panels) and vascular invasion of the outer nuclear layer (white arrows in lower panels), both with associated CD45+ immune cells. ( B ) Cdh5CreER;Tgfbr2 CKO/- retinas showing CNV (white arrows in central panels) and an anastomosis between retinal and choroidal vasculatures (white arrows in lower panels), with intraretinal EC marker CLDN5, choroidal EC marker PLVAP, and pan-vascular marker COL4 (collagen4). ( C ) Upper right panels, in the Cdh5CreER;Tgfbr1 CKO/- retina, CNV (white arrows) is derived from choroidal vasculature, marked by PLVAP. Lower right panels, CNV (white arrows) is present in the subretinal space, i.e., on the retinal side of the RPE, which is marked by RPE65. Abbreviations: Ch., choroid; ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer; RPE, retinal pigment epithelium. The ages of the mice are indicated in postnatal days (P) for this and all other figures. Scale bars, 100 µm.

    Journal: eLife

    Article Title: Vascular endothelial-specific loss of TGF-beta signaling as a model for choroidal neovascularization and central nervous system vascular inflammation

    doi: 10.7554/eLife.107018

    Figure Lengend Snippet: ( A ) Cdh5CreER;Tgfbr1 CKO/- retinas showing CNV (white arrows in central panels) and vascular invasion of the outer nuclear layer (white arrows in lower panels), both with associated CD45+ immune cells. ( B ) Cdh5CreER;Tgfbr2 CKO/- retinas showing CNV (white arrows in central panels) and an anastomosis between retinal and choroidal vasculatures (white arrows in lower panels), with intraretinal EC marker CLDN5, choroidal EC marker PLVAP, and pan-vascular marker COL4 (collagen4). ( C ) Upper right panels, in the Cdh5CreER;Tgfbr1 CKO/- retina, CNV (white arrows) is derived from choroidal vasculature, marked by PLVAP. Lower right panels, CNV (white arrows) is present in the subretinal space, i.e., on the retinal side of the RPE, which is marked by RPE65. Abbreviations: Ch., choroid; ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer; RPE, retinal pigment epithelium. The ages of the mice are indicated in postnatal days (P) for this and all other figures. Scale bars, 100 µm.

    Article Snippet: The following antibodies were used for tissue immunohistochemistry: rat mAb anti-mouse PLVAP/MECA-32 (BD Biosciences 553849); rat mAb anti-mouse CD31 (BD Biosciences 553370); rat anti-mouse ICAM-1 (Invitrogen 14-0542-82); rat mAb anti-mouse F4/80 (Bio-Rad MCA497G); rat mAb anti-mouse CD206 (Bio-Rad MCA2235); rat mAb anti-mouse PU.1/Spi-1 (R&D Systems MAB7124); mouse mAb anti-alpha SMA, Cy3 conjugate (Sigma-Aldrich C6198); mouse mAb anti-CLDN5, Alexa Fluor 488 conjugate (Thermo Fisher Scientific 352588); mouse mAb anti-RPE65, Dylight 550 conjugate (Invitrogen MA5-16044); rabbit polyclonal anti-Collagen IV (Novus Biologicals NB120-6586); rabbit polyclonal anti-NG2 Chondroitin Sulfate Proteoglycan (Millipore AB5320); rabbit mAb anti-ASC/TMS1 (Cell Signaling 67824S); rabbit mAb anti-cleaved Caspase-3 (Cell Signaling 9664S); rabbit mAb anti-HIF-1alpha (Cell Signaling 36169S); rabbit mAb anti-P-SMAD1/5/9 (Cell Signaling 13820S); Armenian hamster mAb anti-CD3e (Invitrogen 14-0031-82); goat polyclonal anti-mouse CD45 (R&D Systems AF114); goat polyclonal anti-Iba1 (Novus Biologicals NB100-1028); chicken polyclonal anti-GFP (Aves Labs GFP-1020); rabbit mAb anti-NFkappaB NF-κB p65 (D14E12; Cell Signaling Technology 8242S); rabbit mAb anti-Integrin alpha 2 (ITGA2; clone GEB, BosterBio M01933); rabbit mAb anti-Integrin alpha 4 (ITGA4; D2E1; Cell Signaling Technology 8440); rabbit mAb anti-TOX/TOX2 (E6G5O; Cell Signaling Technology 36778S).

    Techniques: Marker, Derivative Assay

    ( A ) Phenotypically wild-type (WT) control retina. ( B ) Cdh5CreER;Tgfbr1 CKO/- retina. Eyes from mice at ~P90 were fresh frozen, sectioned, and immunostained. RPE65 marks the retinal pigment epithelium (RPE). In the Cdh5CreER;Tgfbr1 CKO/- retina in ( B ), PECAM1+ ECs and associated RPE cells can be seen protruding into the subretinal space in an ~200 µm wide region (white arrows). ( C ) Quantification of CNV tufts in frozen sections of adult Cdh5CreER;Tgfbr1 CKO/- retinas, Cdh5CreER;Tgfbr2 CKO/- retinas, Ndp KO retinas, and age-matched control retinas. Each data point represents a single whole eye section. CNV tufts were seen only in Cdh5CreER;Tgfbr1 CKO/- retinas. ( D ) Retina flatmounts from Cdh5CreER;Tgfbr1 CKO/- ~P90 mice show scattered CD45+ cells (upper panel) and variably sized zones of CNV with vessel-associated CD45+ cells (lower panel). The stacked Z-planes of the images are at the level of photoreceptors and RPE. Retinas were mounted photoreceptor side up. All other retina flatmount images were mounted ganglion cell side up. The confocal microscope collects images from above. Abbreviations: Ch., choroid; ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer; P, postnatal day. Scale bars, 100 µm.

    Journal: eLife

    Article Title: Vascular endothelial-specific loss of TGF-beta signaling as a model for choroidal neovascularization and central nervous system vascular inflammation

    doi: 10.7554/eLife.107018

    Figure Lengend Snippet: ( A ) Phenotypically wild-type (WT) control retina. ( B ) Cdh5CreER;Tgfbr1 CKO/- retina. Eyes from mice at ~P90 were fresh frozen, sectioned, and immunostained. RPE65 marks the retinal pigment epithelium (RPE). In the Cdh5CreER;Tgfbr1 CKO/- retina in ( B ), PECAM1+ ECs and associated RPE cells can be seen protruding into the subretinal space in an ~200 µm wide region (white arrows). ( C ) Quantification of CNV tufts in frozen sections of adult Cdh5CreER;Tgfbr1 CKO/- retinas, Cdh5CreER;Tgfbr2 CKO/- retinas, Ndp KO retinas, and age-matched control retinas. Each data point represents a single whole eye section. CNV tufts were seen only in Cdh5CreER;Tgfbr1 CKO/- retinas. ( D ) Retina flatmounts from Cdh5CreER;Tgfbr1 CKO/- ~P90 mice show scattered CD45+ cells (upper panel) and variably sized zones of CNV with vessel-associated CD45+ cells (lower panel). The stacked Z-planes of the images are at the level of photoreceptors and RPE. Retinas were mounted photoreceptor side up. All other retina flatmount images were mounted ganglion cell side up. The confocal microscope collects images from above. Abbreviations: Ch., choroid; ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer; P, postnatal day. Scale bars, 100 µm.

    Article Snippet: The following antibodies were used for tissue immunohistochemistry: rat mAb anti-mouse PLVAP/MECA-32 (BD Biosciences 553849); rat mAb anti-mouse CD31 (BD Biosciences 553370); rat anti-mouse ICAM-1 (Invitrogen 14-0542-82); rat mAb anti-mouse F4/80 (Bio-Rad MCA497G); rat mAb anti-mouse CD206 (Bio-Rad MCA2235); rat mAb anti-mouse PU.1/Spi-1 (R&D Systems MAB7124); mouse mAb anti-alpha SMA, Cy3 conjugate (Sigma-Aldrich C6198); mouse mAb anti-CLDN5, Alexa Fluor 488 conjugate (Thermo Fisher Scientific 352588); mouse mAb anti-RPE65, Dylight 550 conjugate (Invitrogen MA5-16044); rabbit polyclonal anti-Collagen IV (Novus Biologicals NB120-6586); rabbit polyclonal anti-NG2 Chondroitin Sulfate Proteoglycan (Millipore AB5320); rabbit mAb anti-ASC/TMS1 (Cell Signaling 67824S); rabbit mAb anti-cleaved Caspase-3 (Cell Signaling 9664S); rabbit mAb anti-HIF-1alpha (Cell Signaling 36169S); rabbit mAb anti-P-SMAD1/5/9 (Cell Signaling 13820S); Armenian hamster mAb anti-CD3e (Invitrogen 14-0031-82); goat polyclonal anti-mouse CD45 (R&D Systems AF114); goat polyclonal anti-Iba1 (Novus Biologicals NB100-1028); chicken polyclonal anti-GFP (Aves Labs GFP-1020); rabbit mAb anti-NFkappaB NF-κB p65 (D14E12; Cell Signaling Technology 8242S); rabbit mAb anti-Integrin alpha 2 (ITGA2; clone GEB, BosterBio M01933); rabbit mAb anti-Integrin alpha 4 (ITGA4; D2E1; Cell Signaling Technology 8440); rabbit mAb anti-TOX/TOX2 (E6G5O; Cell Signaling Technology 36778S).

    Techniques: Control, Microscopy

    Collagen-4 (COL4) marks ECM, including perivascular ECM. ICAM1 is unchanged. P14 corresponds to the peak of CD45+ cell counts in the Cdh5CreER;Tgfbr1 CKO/- retina. Scale bar, 50 µm.

    Journal: eLife

    Article Title: Vascular endothelial-specific loss of TGF-beta signaling as a model for choroidal neovascularization and central nervous system vascular inflammation

    doi: 10.7554/eLife.107018

    Figure Lengend Snippet: Collagen-4 (COL4) marks ECM, including perivascular ECM. ICAM1 is unchanged. P14 corresponds to the peak of CD45+ cell counts in the Cdh5CreER;Tgfbr1 CKO/- retina. Scale bar, 50 µm.

    Article Snippet: The following antibodies were used for tissue immunohistochemistry: rat mAb anti-mouse PLVAP/MECA-32 (BD Biosciences 553849); rat mAb anti-mouse CD31 (BD Biosciences 553370); rat anti-mouse ICAM-1 (Invitrogen 14-0542-82); rat mAb anti-mouse F4/80 (Bio-Rad MCA497G); rat mAb anti-mouse CD206 (Bio-Rad MCA2235); rat mAb anti-mouse PU.1/Spi-1 (R&D Systems MAB7124); mouse mAb anti-alpha SMA, Cy3 conjugate (Sigma-Aldrich C6198); mouse mAb anti-CLDN5, Alexa Fluor 488 conjugate (Thermo Fisher Scientific 352588); mouse mAb anti-RPE65, Dylight 550 conjugate (Invitrogen MA5-16044); rabbit polyclonal anti-Collagen IV (Novus Biologicals NB120-6586); rabbit polyclonal anti-NG2 Chondroitin Sulfate Proteoglycan (Millipore AB5320); rabbit mAb anti-ASC/TMS1 (Cell Signaling 67824S); rabbit mAb anti-cleaved Caspase-3 (Cell Signaling 9664S); rabbit mAb anti-HIF-1alpha (Cell Signaling 36169S); rabbit mAb anti-P-SMAD1/5/9 (Cell Signaling 13820S); Armenian hamster mAb anti-CD3e (Invitrogen 14-0031-82); goat polyclonal anti-mouse CD45 (R&D Systems AF114); goat polyclonal anti-Iba1 (Novus Biologicals NB100-1028); chicken polyclonal anti-GFP (Aves Labs GFP-1020); rabbit mAb anti-NFkappaB NF-κB p65 (D14E12; Cell Signaling Technology 8242S); rabbit mAb anti-Integrin alpha 2 (ITGA2; clone GEB, BosterBio M01933); rabbit mAb anti-Integrin alpha 4 (ITGA4; D2E1; Cell Signaling Technology 8440); rabbit mAb anti-TOX/TOX2 (E6G5O; Cell Signaling Technology 36778S).

    Techniques:

    ( A, B ) Co-localization of the nuclear-localized GFP reporter with the EC transcription factor ERG and non-co-localization with the macrophage marker CD45 in ( A ) a flatmount of choroid and in ( B ) a vibratome section of small intestine. In the rightmost panel, GFP and CD45 are seen to have distinct localizations in both tissues. ( C ) Retina flatmounts showing non-overlapping patterns of GFP (elongated EC nuclei) and CD45 (microglia). Scale bars, 100 µm.

    Journal: eLife

    Article Title: Vascular endothelial-specific loss of TGF-beta signaling as a model for choroidal neovascularization and central nervous system vascular inflammation

    doi: 10.7554/eLife.107018

    Figure Lengend Snippet: ( A, B ) Co-localization of the nuclear-localized GFP reporter with the EC transcription factor ERG and non-co-localization with the macrophage marker CD45 in ( A ) a flatmount of choroid and in ( B ) a vibratome section of small intestine. In the rightmost panel, GFP and CD45 are seen to have distinct localizations in both tissues. ( C ) Retina flatmounts showing non-overlapping patterns of GFP (elongated EC nuclei) and CD45 (microglia). Scale bars, 100 µm.

    Article Snippet: The following antibodies were used for tissue immunohistochemistry: rat mAb anti-mouse PLVAP/MECA-32 (BD Biosciences 553849); rat mAb anti-mouse CD31 (BD Biosciences 553370); rat anti-mouse ICAM-1 (Invitrogen 14-0542-82); rat mAb anti-mouse F4/80 (Bio-Rad MCA497G); rat mAb anti-mouse CD206 (Bio-Rad MCA2235); rat mAb anti-mouse PU.1/Spi-1 (R&D Systems MAB7124); mouse mAb anti-alpha SMA, Cy3 conjugate (Sigma-Aldrich C6198); mouse mAb anti-CLDN5, Alexa Fluor 488 conjugate (Thermo Fisher Scientific 352588); mouse mAb anti-RPE65, Dylight 550 conjugate (Invitrogen MA5-16044); rabbit polyclonal anti-Collagen IV (Novus Biologicals NB120-6586); rabbit polyclonal anti-NG2 Chondroitin Sulfate Proteoglycan (Millipore AB5320); rabbit mAb anti-ASC/TMS1 (Cell Signaling 67824S); rabbit mAb anti-cleaved Caspase-3 (Cell Signaling 9664S); rabbit mAb anti-HIF-1alpha (Cell Signaling 36169S); rabbit mAb anti-P-SMAD1/5/9 (Cell Signaling 13820S); Armenian hamster mAb anti-CD3e (Invitrogen 14-0031-82); goat polyclonal anti-mouse CD45 (R&D Systems AF114); goat polyclonal anti-Iba1 (Novus Biologicals NB100-1028); chicken polyclonal anti-GFP (Aves Labs GFP-1020); rabbit mAb anti-NFkappaB NF-κB p65 (D14E12; Cell Signaling Technology 8242S); rabbit mAb anti-Integrin alpha 2 (ITGA2; clone GEB, BosterBio M01933); rabbit mAb anti-Integrin alpha 4 (ITGA4; D2E1; Cell Signaling Technology 8440); rabbit mAb anti-TOX/TOX2 (E6G5O; Cell Signaling Technology 36778S).

    Techniques: Marker

    A retina flatmount from a P17 Cdh5CreER;Tgfbr1 CKO/- ;Rosa26-LSL-SUN1-sfGFP-6xmyc mouse shows the phenotype of CD45+ immune cell accumulation following loss of TGF-beta signaling. GFP expressed from the LSL reporter co-localizes with ERG and shows no co-localization with CD45. Scale bar, 100 µm.

    Journal: eLife

    Article Title: Vascular endothelial-specific loss of TGF-beta signaling as a model for choroidal neovascularization and central nervous system vascular inflammation

    doi: 10.7554/eLife.107018

    Figure Lengend Snippet: A retina flatmount from a P17 Cdh5CreER;Tgfbr1 CKO/- ;Rosa26-LSL-SUN1-sfGFP-6xmyc mouse shows the phenotype of CD45+ immune cell accumulation following loss of TGF-beta signaling. GFP expressed from the LSL reporter co-localizes with ERG and shows no co-localization with CD45. Scale bar, 100 µm.

    Article Snippet: The following antibodies were used for tissue immunohistochemistry: rat mAb anti-mouse PLVAP/MECA-32 (BD Biosciences 553849); rat mAb anti-mouse CD31 (BD Biosciences 553370); rat anti-mouse ICAM-1 (Invitrogen 14-0542-82); rat mAb anti-mouse F4/80 (Bio-Rad MCA497G); rat mAb anti-mouse CD206 (Bio-Rad MCA2235); rat mAb anti-mouse PU.1/Spi-1 (R&D Systems MAB7124); mouse mAb anti-alpha SMA, Cy3 conjugate (Sigma-Aldrich C6198); mouse mAb anti-CLDN5, Alexa Fluor 488 conjugate (Thermo Fisher Scientific 352588); mouse mAb anti-RPE65, Dylight 550 conjugate (Invitrogen MA5-16044); rabbit polyclonal anti-Collagen IV (Novus Biologicals NB120-6586); rabbit polyclonal anti-NG2 Chondroitin Sulfate Proteoglycan (Millipore AB5320); rabbit mAb anti-ASC/TMS1 (Cell Signaling 67824S); rabbit mAb anti-cleaved Caspase-3 (Cell Signaling 9664S); rabbit mAb anti-HIF-1alpha (Cell Signaling 36169S); rabbit mAb anti-P-SMAD1/5/9 (Cell Signaling 13820S); Armenian hamster mAb anti-CD3e (Invitrogen 14-0031-82); goat polyclonal anti-mouse CD45 (R&D Systems AF114); goat polyclonal anti-Iba1 (Novus Biologicals NB100-1028); chicken polyclonal anti-GFP (Aves Labs GFP-1020); rabbit mAb anti-NFkappaB NF-κB p65 (D14E12; Cell Signaling Technology 8242S); rabbit mAb anti-Integrin alpha 2 (ITGA2; clone GEB, BosterBio M01933); rabbit mAb anti-Integrin alpha 4 (ITGA4; D2E1; Cell Signaling Technology 8440); rabbit mAb anti-TOX/TOX2 (E6G5O; Cell Signaling Technology 36778S).

    Techniques:

    ( A ) Microglia and retinal endothelial cells (ECs) were visualized by immunostaining for CD45 and PECAM1 (upper two panels). By visual inspection, the distributions of the two cell types appear to be uncorrelated. For quantification, microglia were visualized by immunostaining for CD45 (plasma membrane) and ASC1 (nucleus) (lower two panels). The regions in the white squares in the left two panels are shown enlarged in the right two panels. ( B ) Quantification of microglial density in each of the three layers of the inner retina, the regions where microglia are found. Bars represent mean ± standard deviation. RGC, retinal ganglion cell layer; IPL, inner plexiform layer; OPL, outer plexiform layer. Scale bars in ( A ): left, 100 µm, right, 50 µm.

    Journal: eLife

    Article Title: Vascular endothelial-specific loss of TGF-beta signaling as a model for choroidal neovascularization and central nervous system vascular inflammation

    doi: 10.7554/eLife.107018

    Figure Lengend Snippet: ( A ) Microglia and retinal endothelial cells (ECs) were visualized by immunostaining for CD45 and PECAM1 (upper two panels). By visual inspection, the distributions of the two cell types appear to be uncorrelated. For quantification, microglia were visualized by immunostaining for CD45 (plasma membrane) and ASC1 (nucleus) (lower two panels). The regions in the white squares in the left two panels are shown enlarged in the right two panels. ( B ) Quantification of microglial density in each of the three layers of the inner retina, the regions where microglia are found. Bars represent mean ± standard deviation. RGC, retinal ganglion cell layer; IPL, inner plexiform layer; OPL, outer plexiform layer. Scale bars in ( A ): left, 100 µm, right, 50 µm.

    Article Snippet: The following antibodies were used for tissue immunohistochemistry: rat mAb anti-mouse PLVAP/MECA-32 (BD Biosciences 553849); rat mAb anti-mouse CD31 (BD Biosciences 553370); rat anti-mouse ICAM-1 (Invitrogen 14-0542-82); rat mAb anti-mouse F4/80 (Bio-Rad MCA497G); rat mAb anti-mouse CD206 (Bio-Rad MCA2235); rat mAb anti-mouse PU.1/Spi-1 (R&D Systems MAB7124); mouse mAb anti-alpha SMA, Cy3 conjugate (Sigma-Aldrich C6198); mouse mAb anti-CLDN5, Alexa Fluor 488 conjugate (Thermo Fisher Scientific 352588); mouse mAb anti-RPE65, Dylight 550 conjugate (Invitrogen MA5-16044); rabbit polyclonal anti-Collagen IV (Novus Biologicals NB120-6586); rabbit polyclonal anti-NG2 Chondroitin Sulfate Proteoglycan (Millipore AB5320); rabbit mAb anti-ASC/TMS1 (Cell Signaling 67824S); rabbit mAb anti-cleaved Caspase-3 (Cell Signaling 9664S); rabbit mAb anti-HIF-1alpha (Cell Signaling 36169S); rabbit mAb anti-P-SMAD1/5/9 (Cell Signaling 13820S); Armenian hamster mAb anti-CD3e (Invitrogen 14-0031-82); goat polyclonal anti-mouse CD45 (R&D Systems AF114); goat polyclonal anti-Iba1 (Novus Biologicals NB100-1028); chicken polyclonal anti-GFP (Aves Labs GFP-1020); rabbit mAb anti-NFkappaB NF-κB p65 (D14E12; Cell Signaling Technology 8242S); rabbit mAb anti-Integrin alpha 2 (ITGA2; clone GEB, BosterBio M01933); rabbit mAb anti-Integrin alpha 4 (ITGA4; D2E1; Cell Signaling Technology 8440); rabbit mAb anti-TOX/TOX2 (E6G5O; Cell Signaling Technology 36778S).

    Techniques: Immunostaining, Clinical Proteomics, Membrane, Standard Deviation

    ( A ) After removing the retina, choroid flatmounts (sclera, choroidal vasculature, and retinal pigment epithelium [RPE]) were imaged from the RPE side. ( B ) The number of CD45+ cells in choroid flatmounts. Bars represent mean ± standard deviation, and p-values, calculated using the Wilcoxon rank-sum test, are shown as *<0.05, **<0.01, ***<0.001, and ****<0.0001. Scale bar, 100 µm.

    Journal: eLife

    Article Title: Vascular endothelial-specific loss of TGF-beta signaling as a model for choroidal neovascularization and central nervous system vascular inflammation

    doi: 10.7554/eLife.107018

    Figure Lengend Snippet: ( A ) After removing the retina, choroid flatmounts (sclera, choroidal vasculature, and retinal pigment epithelium [RPE]) were imaged from the RPE side. ( B ) The number of CD45+ cells in choroid flatmounts. Bars represent mean ± standard deviation, and p-values, calculated using the Wilcoxon rank-sum test, are shown as *<0.05, **<0.01, ***<0.001, and ****<0.0001. Scale bar, 100 µm.

    Article Snippet: The following antibodies were used for tissue immunohistochemistry: rat mAb anti-mouse PLVAP/MECA-32 (BD Biosciences 553849); rat mAb anti-mouse CD31 (BD Biosciences 553370); rat anti-mouse ICAM-1 (Invitrogen 14-0542-82); rat mAb anti-mouse F4/80 (Bio-Rad MCA497G); rat mAb anti-mouse CD206 (Bio-Rad MCA2235); rat mAb anti-mouse PU.1/Spi-1 (R&D Systems MAB7124); mouse mAb anti-alpha SMA, Cy3 conjugate (Sigma-Aldrich C6198); mouse mAb anti-CLDN5, Alexa Fluor 488 conjugate (Thermo Fisher Scientific 352588); mouse mAb anti-RPE65, Dylight 550 conjugate (Invitrogen MA5-16044); rabbit polyclonal anti-Collagen IV (Novus Biologicals NB120-6586); rabbit polyclonal anti-NG2 Chondroitin Sulfate Proteoglycan (Millipore AB5320); rabbit mAb anti-ASC/TMS1 (Cell Signaling 67824S); rabbit mAb anti-cleaved Caspase-3 (Cell Signaling 9664S); rabbit mAb anti-HIF-1alpha (Cell Signaling 36169S); rabbit mAb anti-P-SMAD1/5/9 (Cell Signaling 13820S); Armenian hamster mAb anti-CD3e (Invitrogen 14-0031-82); goat polyclonal anti-mouse CD45 (R&D Systems AF114); goat polyclonal anti-Iba1 (Novus Biologicals NB100-1028); chicken polyclonal anti-GFP (Aves Labs GFP-1020); rabbit mAb anti-NFkappaB NF-κB p65 (D14E12; Cell Signaling Technology 8242S); rabbit mAb anti-Integrin alpha 2 (ITGA2; clone GEB, BosterBio M01933); rabbit mAb anti-Integrin alpha 4 (ITGA4; D2E1; Cell Signaling Technology 8440); rabbit mAb anti-TOX/TOX2 (E6G5O; Cell Signaling Technology 36778S).

    Techniques: Standard Deviation

    ( A ) Retina flatmounts from mice with the indicated genotypes were immunostained for cleaved Caspase 3, CD45, and PECAM1. In Cdh5CreER;Tgfbr1 CKO/- and Cdh5CreER;Tgfbr1 CKO/- ;Tgfbr2 CKO/- retinas, more than 20% of CD45+ cells are also positive for cleaved caspase 3. ( B ) Quantifying the number of cleaved caspase 3+ cells in control, Fzd4 -/- , and Cdh5CreER;Tgfbr1 CKO/- retinas. Bars represent mean ± standard deviation, and p-values, calculated using the Wilcoxon rank-sum test, are shown as *<0.05, **<0.01, ***<0.001, and ****<0.0001. n.s., not significant. Scale bar, 100 µm.

    Journal: eLife

    Article Title: Vascular endothelial-specific loss of TGF-beta signaling as a model for choroidal neovascularization and central nervous system vascular inflammation

    doi: 10.7554/eLife.107018

    Figure Lengend Snippet: ( A ) Retina flatmounts from mice with the indicated genotypes were immunostained for cleaved Caspase 3, CD45, and PECAM1. In Cdh5CreER;Tgfbr1 CKO/- and Cdh5CreER;Tgfbr1 CKO/- ;Tgfbr2 CKO/- retinas, more than 20% of CD45+ cells are also positive for cleaved caspase 3. ( B ) Quantifying the number of cleaved caspase 3+ cells in control, Fzd4 -/- , and Cdh5CreER;Tgfbr1 CKO/- retinas. Bars represent mean ± standard deviation, and p-values, calculated using the Wilcoxon rank-sum test, are shown as *<0.05, **<0.01, ***<0.001, and ****<0.0001. n.s., not significant. Scale bar, 100 µm.

    Article Snippet: The following antibodies were used for tissue immunohistochemistry: rat mAb anti-mouse PLVAP/MECA-32 (BD Biosciences 553849); rat mAb anti-mouse CD31 (BD Biosciences 553370); rat anti-mouse ICAM-1 (Invitrogen 14-0542-82); rat mAb anti-mouse F4/80 (Bio-Rad MCA497G); rat mAb anti-mouse CD206 (Bio-Rad MCA2235); rat mAb anti-mouse PU.1/Spi-1 (R&D Systems MAB7124); mouse mAb anti-alpha SMA, Cy3 conjugate (Sigma-Aldrich C6198); mouse mAb anti-CLDN5, Alexa Fluor 488 conjugate (Thermo Fisher Scientific 352588); mouse mAb anti-RPE65, Dylight 550 conjugate (Invitrogen MA5-16044); rabbit polyclonal anti-Collagen IV (Novus Biologicals NB120-6586); rabbit polyclonal anti-NG2 Chondroitin Sulfate Proteoglycan (Millipore AB5320); rabbit mAb anti-ASC/TMS1 (Cell Signaling 67824S); rabbit mAb anti-cleaved Caspase-3 (Cell Signaling 9664S); rabbit mAb anti-HIF-1alpha (Cell Signaling 36169S); rabbit mAb anti-P-SMAD1/5/9 (Cell Signaling 13820S); Armenian hamster mAb anti-CD3e (Invitrogen 14-0031-82); goat polyclonal anti-mouse CD45 (R&D Systems AF114); goat polyclonal anti-Iba1 (Novus Biologicals NB100-1028); chicken polyclonal anti-GFP (Aves Labs GFP-1020); rabbit mAb anti-NFkappaB NF-κB p65 (D14E12; Cell Signaling Technology 8242S); rabbit mAb anti-Integrin alpha 2 (ITGA2; clone GEB, BosterBio M01933); rabbit mAb anti-Integrin alpha 4 (ITGA4; D2E1; Cell Signaling Technology 8440); rabbit mAb anti-TOX/TOX2 (E6G5O; Cell Signaling Technology 36778S).

    Techniques: Control, Standard Deviation

    ( A ) Sections of control and Vsx2-Cre;Vegfa CKO/CKO retinas immunostained with anti-PECAM1 to visualize the vasculature. ( B ) False color images of control and Vsx2-Cre;Vegfa CKO/CKO flatmount retinas showing a stacked Z-series color-coded by the depth of PECAM1 immunostained vasculature. Blue, vitreal surface; green, inner plexiform layer; red, outer plexiform layer. ( C ) Control and Vsx2-Cre;Vegfa CKO/CKO flatmount retinas immunostained with anti-PECAM1. ( D ) Control and Vsx2-Cre;Vegfa CKO/CKO flatmount retinas immunostained for ASC and CD45. ( E ) Quantification of CD45+/ASC+ cells. Scale bars, ( A ), ( B ), and ( D ), 100 µm; ( C ) 500 µm.

    Journal: eLife

    Article Title: Vascular endothelial-specific loss of TGF-beta signaling as a model for choroidal neovascularization and central nervous system vascular inflammation

    doi: 10.7554/eLife.107018

    Figure Lengend Snippet: ( A ) Sections of control and Vsx2-Cre;Vegfa CKO/CKO retinas immunostained with anti-PECAM1 to visualize the vasculature. ( B ) False color images of control and Vsx2-Cre;Vegfa CKO/CKO flatmount retinas showing a stacked Z-series color-coded by the depth of PECAM1 immunostained vasculature. Blue, vitreal surface; green, inner plexiform layer; red, outer plexiform layer. ( C ) Control and Vsx2-Cre;Vegfa CKO/CKO flatmount retinas immunostained with anti-PECAM1. ( D ) Control and Vsx2-Cre;Vegfa CKO/CKO flatmount retinas immunostained for ASC and CD45. ( E ) Quantification of CD45+/ASC+ cells. Scale bars, ( A ), ( B ), and ( D ), 100 µm; ( C ) 500 µm.

    Article Snippet: The following antibodies were used for tissue immunohistochemistry: rat mAb anti-mouse PLVAP/MECA-32 (BD Biosciences 553849); rat mAb anti-mouse CD31 (BD Biosciences 553370); rat anti-mouse ICAM-1 (Invitrogen 14-0542-82); rat mAb anti-mouse F4/80 (Bio-Rad MCA497G); rat mAb anti-mouse CD206 (Bio-Rad MCA2235); rat mAb anti-mouse PU.1/Spi-1 (R&D Systems MAB7124); mouse mAb anti-alpha SMA, Cy3 conjugate (Sigma-Aldrich C6198); mouse mAb anti-CLDN5, Alexa Fluor 488 conjugate (Thermo Fisher Scientific 352588); mouse mAb anti-RPE65, Dylight 550 conjugate (Invitrogen MA5-16044); rabbit polyclonal anti-Collagen IV (Novus Biologicals NB120-6586); rabbit polyclonal anti-NG2 Chondroitin Sulfate Proteoglycan (Millipore AB5320); rabbit mAb anti-ASC/TMS1 (Cell Signaling 67824S); rabbit mAb anti-cleaved Caspase-3 (Cell Signaling 9664S); rabbit mAb anti-HIF-1alpha (Cell Signaling 36169S); rabbit mAb anti-P-SMAD1/5/9 (Cell Signaling 13820S); Armenian hamster mAb anti-CD3e (Invitrogen 14-0031-82); goat polyclonal anti-mouse CD45 (R&D Systems AF114); goat polyclonal anti-Iba1 (Novus Biologicals NB100-1028); chicken polyclonal anti-GFP (Aves Labs GFP-1020); rabbit mAb anti-NFkappaB NF-κB p65 (D14E12; Cell Signaling Technology 8242S); rabbit mAb anti-Integrin alpha 2 (ITGA2; clone GEB, BosterBio M01933); rabbit mAb anti-Integrin alpha 4 (ITGA4; D2E1; Cell Signaling Technology 8440); rabbit mAb anti-TOX/TOX2 (E6G5O; Cell Signaling Technology 36778S).

    Techniques: Control

    ( A ) Upper six panels show a control retina flatmount. Lower six panels show a Cdh5CreER;Tgfbr1 CKO/+ ;Tgfbr2 CKO/- retina flatmount. ASC and CD45 label immune cells, including microglia. The Z-plane is indicated by the numbers at the bottom of each image. The nerve fiber layer (Z-plane 3) and the inner plexiform layer (Z-planes 10–11) are shown schematically in ( B ) and ( C ). ( B and C ) Retinal schematics showing the relationship of the vasculature and the three retinal layers. Confocal Z-planes are numbered at right. ( D ) Immune cells and venous ECs in control and Cdh5CreER;Tgfbr1 CKO/- retinas. In the lower image, three of the ‘impressions’ of CD45+ immune in the distribution of PECAM1 on the EC surface are highlighted with white arrows. A, artery; V, vein. NFL, nerve fiber layer; GCL, ganglion cell layer; IPL, inner plexiform layer; INL, inner nuclear layer; OPL, outer plexiform layer; ONL, outer nuclear layer; IS/OS, inner segment/outer segment. Scale bars in ( A ), 100 µm. Scale bar in ( D ), 50 µm.

    Journal: eLife

    Article Title: Vascular endothelial-specific loss of TGF-beta signaling as a model for choroidal neovascularization and central nervous system vascular inflammation

    doi: 10.7554/eLife.107018

    Figure Lengend Snippet: ( A ) Upper six panels show a control retina flatmount. Lower six panels show a Cdh5CreER;Tgfbr1 CKO/+ ;Tgfbr2 CKO/- retina flatmount. ASC and CD45 label immune cells, including microglia. The Z-plane is indicated by the numbers at the bottom of each image. The nerve fiber layer (Z-plane 3) and the inner plexiform layer (Z-planes 10–11) are shown schematically in ( B ) and ( C ). ( B and C ) Retinal schematics showing the relationship of the vasculature and the three retinal layers. Confocal Z-planes are numbered at right. ( D ) Immune cells and venous ECs in control and Cdh5CreER;Tgfbr1 CKO/- retinas. In the lower image, three of the ‘impressions’ of CD45+ immune in the distribution of PECAM1 on the EC surface are highlighted with white arrows. A, artery; V, vein. NFL, nerve fiber layer; GCL, ganglion cell layer; IPL, inner plexiform layer; INL, inner nuclear layer; OPL, outer plexiform layer; ONL, outer nuclear layer; IS/OS, inner segment/outer segment. Scale bars in ( A ), 100 µm. Scale bar in ( D ), 50 µm.

    Article Snippet: The following antibodies were used for tissue immunohistochemistry: rat mAb anti-mouse PLVAP/MECA-32 (BD Biosciences 553849); rat mAb anti-mouse CD31 (BD Biosciences 553370); rat anti-mouse ICAM-1 (Invitrogen 14-0542-82); rat mAb anti-mouse F4/80 (Bio-Rad MCA497G); rat mAb anti-mouse CD206 (Bio-Rad MCA2235); rat mAb anti-mouse PU.1/Spi-1 (R&D Systems MAB7124); mouse mAb anti-alpha SMA, Cy3 conjugate (Sigma-Aldrich C6198); mouse mAb anti-CLDN5, Alexa Fluor 488 conjugate (Thermo Fisher Scientific 352588); mouse mAb anti-RPE65, Dylight 550 conjugate (Invitrogen MA5-16044); rabbit polyclonal anti-Collagen IV (Novus Biologicals NB120-6586); rabbit polyclonal anti-NG2 Chondroitin Sulfate Proteoglycan (Millipore AB5320); rabbit mAb anti-ASC/TMS1 (Cell Signaling 67824S); rabbit mAb anti-cleaved Caspase-3 (Cell Signaling 9664S); rabbit mAb anti-HIF-1alpha (Cell Signaling 36169S); rabbit mAb anti-P-SMAD1/5/9 (Cell Signaling 13820S); Armenian hamster mAb anti-CD3e (Invitrogen 14-0031-82); goat polyclonal anti-mouse CD45 (R&D Systems AF114); goat polyclonal anti-Iba1 (Novus Biologicals NB100-1028); chicken polyclonal anti-GFP (Aves Labs GFP-1020); rabbit mAb anti-NFkappaB NF-κB p65 (D14E12; Cell Signaling Technology 8242S); rabbit mAb anti-Integrin alpha 2 (ITGA2; clone GEB, BosterBio M01933); rabbit mAb anti-Integrin alpha 4 (ITGA4; D2E1; Cell Signaling Technology 8440); rabbit mAb anti-TOX/TOX2 (E6G5O; Cell Signaling Technology 36778S).

    Techniques: Control

    ( A ) Upper three panels show a control retina flatmount with sparse CD45+ microglia and PECAM1+ vasculature, imaged at three Z-planes (indicated by the numbers in the lower left corner of each image) corresponding to the NFL, IPL, and OPL. The lower nine panels show a series of Z-planes through the full thickness of a Cdh5CreER;Tgfbr1 CKO/- retina flatmount with many CD45+ immune cells in contact with the vasculature. ( B and C ) Retinal schematics showing the relationship of the vasculature and the three retinal layers. Confocal Z-planes are numbered at right. NFL, nerve fiber layer; GCL, ganglion cell layer; IPL, inner plexiform layer; INL, inner nuclear layer; OPL, outer plexiform layer; ONL, outer nuclear layer; IS/OS, inner segment/outer segment. Scale bars in ( A ), 100 µm.

    Journal: eLife

    Article Title: Vascular endothelial-specific loss of TGF-beta signaling as a model for choroidal neovascularization and central nervous system vascular inflammation

    doi: 10.7554/eLife.107018

    Figure Lengend Snippet: ( A ) Upper three panels show a control retina flatmount with sparse CD45+ microglia and PECAM1+ vasculature, imaged at three Z-planes (indicated by the numbers in the lower left corner of each image) corresponding to the NFL, IPL, and OPL. The lower nine panels show a series of Z-planes through the full thickness of a Cdh5CreER;Tgfbr1 CKO/- retina flatmount with many CD45+ immune cells in contact with the vasculature. ( B and C ) Retinal schematics showing the relationship of the vasculature and the three retinal layers. Confocal Z-planes are numbered at right. NFL, nerve fiber layer; GCL, ganglion cell layer; IPL, inner plexiform layer; INL, inner nuclear layer; OPL, outer plexiform layer; ONL, outer nuclear layer; IS/OS, inner segment/outer segment. Scale bars in ( A ), 100 µm.

    Article Snippet: The following antibodies were used for tissue immunohistochemistry: rat mAb anti-mouse PLVAP/MECA-32 (BD Biosciences 553849); rat mAb anti-mouse CD31 (BD Biosciences 553370); rat anti-mouse ICAM-1 (Invitrogen 14-0542-82); rat mAb anti-mouse F4/80 (Bio-Rad MCA497G); rat mAb anti-mouse CD206 (Bio-Rad MCA2235); rat mAb anti-mouse PU.1/Spi-1 (R&D Systems MAB7124); mouse mAb anti-alpha SMA, Cy3 conjugate (Sigma-Aldrich C6198); mouse mAb anti-CLDN5, Alexa Fluor 488 conjugate (Thermo Fisher Scientific 352588); mouse mAb anti-RPE65, Dylight 550 conjugate (Invitrogen MA5-16044); rabbit polyclonal anti-Collagen IV (Novus Biologicals NB120-6586); rabbit polyclonal anti-NG2 Chondroitin Sulfate Proteoglycan (Millipore AB5320); rabbit mAb anti-ASC/TMS1 (Cell Signaling 67824S); rabbit mAb anti-cleaved Caspase-3 (Cell Signaling 9664S); rabbit mAb anti-HIF-1alpha (Cell Signaling 36169S); rabbit mAb anti-P-SMAD1/5/9 (Cell Signaling 13820S); Armenian hamster mAb anti-CD3e (Invitrogen 14-0031-82); goat polyclonal anti-mouse CD45 (R&D Systems AF114); goat polyclonal anti-Iba1 (Novus Biologicals NB100-1028); chicken polyclonal anti-GFP (Aves Labs GFP-1020); rabbit mAb anti-NFkappaB NF-κB p65 (D14E12; Cell Signaling Technology 8242S); rabbit mAb anti-Integrin alpha 2 (ITGA2; clone GEB, BosterBio M01933); rabbit mAb anti-Integrin alpha 4 (ITGA4; D2E1; Cell Signaling Technology 8440); rabbit mAb anti-TOX/TOX2 (E6G5O; Cell Signaling Technology 36778S).

    Techniques: Control

    ( A ) Endogenous IgG in control and Cdh5CreER;Tgfbr1 CKO/- brains at postnatal day (P)14 and P24. IgG accumulation is minimal in control brains at P14 and P24 but is readily detectable in Cdh5CreER;Tgfbr1 CKO/- brains at P14 but not at P24. ( B ) ECs (visualized with PECAM1) and pericytes (visualized with NG2) in control and Cdh5CreER;Tgfbr1 CKO/- brains at P14. SMA immunostaining visualizes arterioles (continuous staining) and veins (patchy staining) in control and Cdh5CreER;Tgfbr1 CKO/- brains, and a subset of capillary-associated pericytes in Cdh5CreER;Tgfbr1 CKO/- brains. ( C ) Immune cells (CD45+ and F4-80+) in control and Cdh5CreER;Tgfbr1 CKO/- brains at P35. Control brains have minimal numbers of immune cells other than resident microglia. Cdh5CreER;Tgfbr1 CKO/- brains show localized regions with concentrated accumulations of immune cells. White squares marked with letters in the sagittal brain images (upper) are enlarged below. Scale bars, 1 mm for whole brain images and 200 µm for enlarged images.

    Journal: eLife

    Article Title: Vascular endothelial-specific loss of TGF-beta signaling as a model for choroidal neovascularization and central nervous system vascular inflammation

    doi: 10.7554/eLife.107018

    Figure Lengend Snippet: ( A ) Endogenous IgG in control and Cdh5CreER;Tgfbr1 CKO/- brains at postnatal day (P)14 and P24. IgG accumulation is minimal in control brains at P14 and P24 but is readily detectable in Cdh5CreER;Tgfbr1 CKO/- brains at P14 but not at P24. ( B ) ECs (visualized with PECAM1) and pericytes (visualized with NG2) in control and Cdh5CreER;Tgfbr1 CKO/- brains at P14. SMA immunostaining visualizes arterioles (continuous staining) and veins (patchy staining) in control and Cdh5CreER;Tgfbr1 CKO/- brains, and a subset of capillary-associated pericytes in Cdh5CreER;Tgfbr1 CKO/- brains. ( C ) Immune cells (CD45+ and F4-80+) in control and Cdh5CreER;Tgfbr1 CKO/- brains at P35. Control brains have minimal numbers of immune cells other than resident microglia. Cdh5CreER;Tgfbr1 CKO/- brains show localized regions with concentrated accumulations of immune cells. White squares marked with letters in the sagittal brain images (upper) are enlarged below. Scale bars, 1 mm for whole brain images and 200 µm for enlarged images.

    Article Snippet: The following antibodies were used for tissue immunohistochemistry: rat mAb anti-mouse PLVAP/MECA-32 (BD Biosciences 553849); rat mAb anti-mouse CD31 (BD Biosciences 553370); rat anti-mouse ICAM-1 (Invitrogen 14-0542-82); rat mAb anti-mouse F4/80 (Bio-Rad MCA497G); rat mAb anti-mouse CD206 (Bio-Rad MCA2235); rat mAb anti-mouse PU.1/Spi-1 (R&D Systems MAB7124); mouse mAb anti-alpha SMA, Cy3 conjugate (Sigma-Aldrich C6198); mouse mAb anti-CLDN5, Alexa Fluor 488 conjugate (Thermo Fisher Scientific 352588); mouse mAb anti-RPE65, Dylight 550 conjugate (Invitrogen MA5-16044); rabbit polyclonal anti-Collagen IV (Novus Biologicals NB120-6586); rabbit polyclonal anti-NG2 Chondroitin Sulfate Proteoglycan (Millipore AB5320); rabbit mAb anti-ASC/TMS1 (Cell Signaling 67824S); rabbit mAb anti-cleaved Caspase-3 (Cell Signaling 9664S); rabbit mAb anti-HIF-1alpha (Cell Signaling 36169S); rabbit mAb anti-P-SMAD1/5/9 (Cell Signaling 13820S); Armenian hamster mAb anti-CD3e (Invitrogen 14-0031-82); goat polyclonal anti-mouse CD45 (R&D Systems AF114); goat polyclonal anti-Iba1 (Novus Biologicals NB100-1028); chicken polyclonal anti-GFP (Aves Labs GFP-1020); rabbit mAb anti-NFkappaB NF-κB p65 (D14E12; Cell Signaling Technology 8242S); rabbit mAb anti-Integrin alpha 2 (ITGA2; clone GEB, BosterBio M01933); rabbit mAb anti-Integrin alpha 4 (ITGA4; D2E1; Cell Signaling Technology 8440); rabbit mAb anti-TOX/TOX2 (E6G5O; Cell Signaling Technology 36778S).

    Techniques: Control, Immunostaining, Staining

    Pharmacological inhibition of IL12β attenuated TAC-induced LV inflammation. (A, B) Representative images and quantified data of LV CD45 + leukocytes performed by immuno-histological staining. (C) Flow cytometry plot for the identification of CD45 + leukocytes in the LV. (D) Quantified data of the number of CD45 + leukocytes per LV. (E) The percentage of immune cell subsets within CD45 + leukocytes. (F) Quantified data of number of different immune cell subsets per LV. *p<0.05; # p<0.05 IgG-treated TAC mice compared with the control; † p<0.05 anti-IL12β-treated TAC mice compared with IgG-treated TAC mice; Neutro, Neutrophils; Mφ, Macrophage; Mono, Monocytes; DCs, Dendritic Cells; NK, Natural Killer Cells; n = 3–5 mice per group.

    Journal: Frontiers in Immunology

    Article Title: Pharmacological inhibition of IL12β is effective in treating pressure overload-induced cardiac inflammation and heart failure

    doi: 10.3389/fimmu.2025.1624940

    Figure Lengend Snippet: Pharmacological inhibition of IL12β attenuated TAC-induced LV inflammation. (A, B) Representative images and quantified data of LV CD45 + leukocytes performed by immuno-histological staining. (C) Flow cytometry plot for the identification of CD45 + leukocytes in the LV. (D) Quantified data of the number of CD45 + leukocytes per LV. (E) The percentage of immune cell subsets within CD45 + leukocytes. (F) Quantified data of number of different immune cell subsets per LV. *p<0.05; # p<0.05 IgG-treated TAC mice compared with the control; † p<0.05 anti-IL12β-treated TAC mice compared with IgG-treated TAC mice; Neutro, Neutrophils; Mφ, Macrophage; Mono, Monocytes; DCs, Dendritic Cells; NK, Natural Killer Cells; n = 3–5 mice per group.

    Article Snippet: The infiltrated CD45 + leukocytes in the LV and lung tissues were stained for visualization with goat anti-CD45 antibody (R&D Systems, AF114, 1:100 dilution) and Alexa Flour-555 conjugated donkey anti-goat secondary antibody (Invitrogen, A21432, 1:1000 dilution).

    Techniques: Inhibition, Staining, Flow Cytometry, Control

    Pharmacological inhibition of IL12β attenuated TAC-induced pulmonary inflammation. (A, B) Representative images and quantified data of infiltrated CD45 + leukocytes in the lung performed by immuno-histological staining. (C) The percentage of immune cell subsets within CD45 + leukocytes. (D) The percentage of different macrophage subsets within F4/80 macrophages. (E) The percentage of immune cell subsets within CD3 + T cells. *p<0.05; # p<0.05 IgG-treated TAC mice compared with the control; † p<0.05 anti-IL12β-treated TAC mice compared with IgG-treated TAC mice; $ p<0.05 anti-IL12β-treated TAC mice compared with the control; Mφ, Macrophage; DCs, Dendritic Cells; NK, Natural Killer Cells; AMφ, Alveolar Mφ; IMφ, Interstitial Mφ; MdMφ, Monocyte-derived Mφ; NKT, Natural Killer T Cells; n = 4–5 mice per group.

    Journal: Frontiers in Immunology

    Article Title: Pharmacological inhibition of IL12β is effective in treating pressure overload-induced cardiac inflammation and heart failure

    doi: 10.3389/fimmu.2025.1624940

    Figure Lengend Snippet: Pharmacological inhibition of IL12β attenuated TAC-induced pulmonary inflammation. (A, B) Representative images and quantified data of infiltrated CD45 + leukocytes in the lung performed by immuno-histological staining. (C) The percentage of immune cell subsets within CD45 + leukocytes. (D) The percentage of different macrophage subsets within F4/80 macrophages. (E) The percentage of immune cell subsets within CD3 + T cells. *p<0.05; # p<0.05 IgG-treated TAC mice compared with the control; † p<0.05 anti-IL12β-treated TAC mice compared with IgG-treated TAC mice; $ p<0.05 anti-IL12β-treated TAC mice compared with the control; Mφ, Macrophage; DCs, Dendritic Cells; NK, Natural Killer Cells; AMφ, Alveolar Mφ; IMφ, Interstitial Mφ; MdMφ, Monocyte-derived Mφ; NKT, Natural Killer T Cells; n = 4–5 mice per group.

    Article Snippet: The infiltrated CD45 + leukocytes in the LV and lung tissues were stained for visualization with goat anti-CD45 antibody (R&D Systems, AF114, 1:100 dilution) and Alexa Flour-555 conjugated donkey anti-goat secondary antibody (Invitrogen, A21432, 1:1000 dilution).

    Techniques: Inhibition, Staining, Control, Derivative Assay

    Anti-IL12β antibody treatment attenuated TAC-induced pulmonary F4/80 + macrophage accumulation and activation. (A) Flow cytometry plots for F4/80 + macrophages. (B) Quantified data of the percentage of F4/80 + cells within CD45 + cells. (C) Flow cytometry plots for the detection of MHCII expression in F4/80 + cells. (D, E) Quantified data of the percentage of MHCII high F4/80 + cells within F4/80 + and CD45 + cells, respectively. (F) Quantified data of mean fluorescent intensity of MHCII in F4/80 + cells. (G) Representative histograms of MHCII expression in F4/80 + cells of the indicated groups. *p<0.05; n = 4–5 mice per group.

    Journal: Frontiers in Immunology

    Article Title: Pharmacological inhibition of IL12β is effective in treating pressure overload-induced cardiac inflammation and heart failure

    doi: 10.3389/fimmu.2025.1624940

    Figure Lengend Snippet: Anti-IL12β antibody treatment attenuated TAC-induced pulmonary F4/80 + macrophage accumulation and activation. (A) Flow cytometry plots for F4/80 + macrophages. (B) Quantified data of the percentage of F4/80 + cells within CD45 + cells. (C) Flow cytometry plots for the detection of MHCII expression in F4/80 + cells. (D, E) Quantified data of the percentage of MHCII high F4/80 + cells within F4/80 + and CD45 + cells, respectively. (F) Quantified data of mean fluorescent intensity of MHCII in F4/80 + cells. (G) Representative histograms of MHCII expression in F4/80 + cells of the indicated groups. *p<0.05; n = 4–5 mice per group.

    Article Snippet: The infiltrated CD45 + leukocytes in the LV and lung tissues were stained for visualization with goat anti-CD45 antibody (R&D Systems, AF114, 1:100 dilution) and Alexa Flour-555 conjugated donkey anti-goat secondary antibody (Invitrogen, A21432, 1:1000 dilution).

    Techniques: Activation Assay, Flow Cytometry, Expressing

    Anti-IL12β antibody treatment attenuated TAC-induced alveolar macrophage accumulation and activation of alveolar and interstitial macrophages. (A) Flow cytometry plots of lung alveolar macrophages (AMφ). (B) Quantified data of the percentage of AMφ, Ly6C low interstitial macrophages (IMφ), and monocyte-derived Ly6C high interstitial macrophages (MdMφ) within CD45 + cells. (C) Flow cytometry plots for the identification of IMφ and MdMφ. (D) Quantified data of the percentage of MHCII high AMφ, MHCII high IMφ, and MHCII high MdMφ within CD45 + leukocytes. (E) Flow cytometry plots for the detection of MHCII expression in AMφ. (F) Quantified data of mean fluorescent intensity of MHCII in AMφ, IMφ, and MdMφ. (G-I) Quantified data of MHCII high AMφ, MHCII high IMφ, and MHCII high MdMφ within AMφ, IMφ, and MdMφ, respectively. (J) Representative histograms of MHCII expression in AMφ, IMφ, and MdMφ of the indicated groups. *p<0.05; # p<0.05 IgG-treated TAC mice compared with the control; † p<0.05 anti-IL12β-treated TAC mice compared with IgG-treated TAC mice; $ p<0.05 anti-IL12β-treated TAC mice compared with the control; MHCII high (MHCII + ); n = 4–5 mice per group.

    Journal: Frontiers in Immunology

    Article Title: Pharmacological inhibition of IL12β is effective in treating pressure overload-induced cardiac inflammation and heart failure

    doi: 10.3389/fimmu.2025.1624940

    Figure Lengend Snippet: Anti-IL12β antibody treatment attenuated TAC-induced alveolar macrophage accumulation and activation of alveolar and interstitial macrophages. (A) Flow cytometry plots of lung alveolar macrophages (AMφ). (B) Quantified data of the percentage of AMφ, Ly6C low interstitial macrophages (IMφ), and monocyte-derived Ly6C high interstitial macrophages (MdMφ) within CD45 + cells. (C) Flow cytometry plots for the identification of IMφ and MdMφ. (D) Quantified data of the percentage of MHCII high AMφ, MHCII high IMφ, and MHCII high MdMφ within CD45 + leukocytes. (E) Flow cytometry plots for the detection of MHCII expression in AMφ. (F) Quantified data of mean fluorescent intensity of MHCII in AMφ, IMφ, and MdMφ. (G-I) Quantified data of MHCII high AMφ, MHCII high IMφ, and MHCII high MdMφ within AMφ, IMφ, and MdMφ, respectively. (J) Representative histograms of MHCII expression in AMφ, IMφ, and MdMφ of the indicated groups. *p<0.05; # p<0.05 IgG-treated TAC mice compared with the control; † p<0.05 anti-IL12β-treated TAC mice compared with IgG-treated TAC mice; $ p<0.05 anti-IL12β-treated TAC mice compared with the control; MHCII high (MHCII + ); n = 4–5 mice per group.

    Article Snippet: The infiltrated CD45 + leukocytes in the LV and lung tissues were stained for visualization with goat anti-CD45 antibody (R&D Systems, AF114, 1:100 dilution) and Alexa Flour-555 conjugated donkey anti-goat secondary antibody (Invitrogen, A21432, 1:1000 dilution).

    Techniques: Activation Assay, Flow Cytometry, Derivative Assay, Expressing, Control

    Anti-IL12β antibody treatment attenuated TAC-induced activation of pulmonary dendritic cells (CD11c high F4/80 - ). (A) Flow cytometry plots of lung CD11c high F4/80 - cells. (B) Quantified data of the percentage of CD11c high F4/80 - cells within CD45 + cells. (C) Flow cytometry plots for the detection of MHCII expression in CD11c high F4/80 - cells. (D, E) Quantified data of the percentage of MHCII high CD11c high F4/80 - cells within CD11c high F4/80 - cells and CD45 + cells, respectively. (F) Quantified data of mean fluorescent intensity of MHCII in CD11c high F4/80 - cells. (G) Representative histograms of MHCII expression in CD11c high F4/80 - cells of the indicated groups. *p<0.05; n = 4–5 mice per group.

    Journal: Frontiers in Immunology

    Article Title: Pharmacological inhibition of IL12β is effective in treating pressure overload-induced cardiac inflammation and heart failure

    doi: 10.3389/fimmu.2025.1624940

    Figure Lengend Snippet: Anti-IL12β antibody treatment attenuated TAC-induced activation of pulmonary dendritic cells (CD11c high F4/80 - ). (A) Flow cytometry plots of lung CD11c high F4/80 - cells. (B) Quantified data of the percentage of CD11c high F4/80 - cells within CD45 + cells. (C) Flow cytometry plots for the detection of MHCII expression in CD11c high F4/80 - cells. (D, E) Quantified data of the percentage of MHCII high CD11c high F4/80 - cells within CD11c high F4/80 - cells and CD45 + cells, respectively. (F) Quantified data of mean fluorescent intensity of MHCII in CD11c high F4/80 - cells. (G) Representative histograms of MHCII expression in CD11c high F4/80 - cells of the indicated groups. *p<0.05; n = 4–5 mice per group.

    Article Snippet: The infiltrated CD45 + leukocytes in the LV and lung tissues were stained for visualization with goat anti-CD45 antibody (R&D Systems, AF114, 1:100 dilution) and Alexa Flour-555 conjugated donkey anti-goat secondary antibody (Invitrogen, A21432, 1:1000 dilution).

    Techniques: Activation Assay, Flow Cytometry, Expressing