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human mouse igf 1r  (R&D Systems)


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    R&D Systems human mouse igf 1r
    Effects of rexinoid treatment on macrophage gene and protein expression in the desiccation stress dry eye model and potential macrophage-derived <t>IGF-1/IGF-1R</t> signaling axis. ( A ) Heatmaps showing z -score–scaled expression of selected latent-time–associated genes in four treatment groups (NS, None [untreated], Veh, and NEt-3IB) in conjunctival monocyte/macrophage lineage cells sorted from CD45⁺ cells after 5 days of desiccating stress (DS5) ( P adj < 0.001). Genes are organized into five functional categories. The color scale represents z -scores of normalized expression ( blue , low; red , high). These data show desiccating stress–associated shifts in macrophage gene programs and their modulation by NEt-3IB, including enrichment of reparative and growth factor–related genes such as Igf1 . Full gene names for the abbreviations are provided in . ( B ) Representative flow cytometry histograms and quantification validating selected macrophage-associated markers in conjunctival immune cells. ( Left ) Representative histograms of CX3CR1 staining with fluorescence-minus-one (FMO) control, with corresponding quantification of the percentage of CD45⁺CD11b⁺CX3CR1⁺ cells. ( Right ) Representative histograms of IGF-1 staining with FMO control, with corresponding quantification of the percentage of CD45⁺CD11b⁺Mrc1⁺IGF-1⁺ cells. Compared with DS5 no treatment and DS5+vehicle controls, DS5+NEt-3IB increased the proportion ofx CX3CR1⁺ and Mrc1⁺IGF-1⁺ myeloid cells. Each dot represents one biological replicate; bars show mean ± SEM. Statistical significance is indicated as shown: ** P < 0.01; **** P < 0.001; **** P < 0.0001; ns, not significant. ( C ) Representative immunofluorescence images showing IGF-1R/WGA/DAPI staining of wholemount conjunctiva showing surface view ( top ) and βIII-tubulin/IGF-1R/DAPI staining in the cornea ( bottom ). IGF-1R localization is shown because IGF-1 is produced by ocular surface resident macrophages, suggesting a potential macrophage-derived IGF-1/IGF-1R signaling axis acting on ocular surface epithelial and neural compartments during DS. Scale bar : 100 µm.
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    Images

    1) Product Images from "Rexinoid NEt-3IB Promotes Resident Macrophage Gene Expression and Mitigates Desiccation-Induced Ocular Surface Disease"

    Article Title: Rexinoid NEt-3IB Promotes Resident Macrophage Gene Expression and Mitigates Desiccation-Induced Ocular Surface Disease

    Journal: Investigative Ophthalmology & Visual Science

    doi: 10.1167/iovs.67.4.31

    Effects of rexinoid treatment on macrophage gene and protein expression in the desiccation stress dry eye model and potential macrophage-derived IGF-1/IGF-1R signaling axis. ( A ) Heatmaps showing z -score–scaled expression of selected latent-time–associated genes in four treatment groups (NS, None [untreated], Veh, and NEt-3IB) in conjunctival monocyte/macrophage lineage cells sorted from CD45⁺ cells after 5 days of desiccating stress (DS5) ( P adj < 0.001). Genes are organized into five functional categories. The color scale represents z -scores of normalized expression ( blue , low; red , high). These data show desiccating stress–associated shifts in macrophage gene programs and their modulation by NEt-3IB, including enrichment of reparative and growth factor–related genes such as Igf1 . Full gene names for the abbreviations are provided in . ( B ) Representative flow cytometry histograms and quantification validating selected macrophage-associated markers in conjunctival immune cells. ( Left ) Representative histograms of CX3CR1 staining with fluorescence-minus-one (FMO) control, with corresponding quantification of the percentage of CD45⁺CD11b⁺CX3CR1⁺ cells. ( Right ) Representative histograms of IGF-1 staining with FMO control, with corresponding quantification of the percentage of CD45⁺CD11b⁺Mrc1⁺IGF-1⁺ cells. Compared with DS5 no treatment and DS5+vehicle controls, DS5+NEt-3IB increased the proportion ofx CX3CR1⁺ and Mrc1⁺IGF-1⁺ myeloid cells. Each dot represents one biological replicate; bars show mean ± SEM. Statistical significance is indicated as shown: ** P < 0.01; **** P < 0.001; **** P < 0.0001; ns, not significant. ( C ) Representative immunofluorescence images showing IGF-1R/WGA/DAPI staining of wholemount conjunctiva showing surface view ( top ) and βIII-tubulin/IGF-1R/DAPI staining in the cornea ( bottom ). IGF-1R localization is shown because IGF-1 is produced by ocular surface resident macrophages, suggesting a potential macrophage-derived IGF-1/IGF-1R signaling axis acting on ocular surface epithelial and neural compartments during DS. Scale bar : 100 µm.
    Figure Legend Snippet: Effects of rexinoid treatment on macrophage gene and protein expression in the desiccation stress dry eye model and potential macrophage-derived IGF-1/IGF-1R signaling axis. ( A ) Heatmaps showing z -score–scaled expression of selected latent-time–associated genes in four treatment groups (NS, None [untreated], Veh, and NEt-3IB) in conjunctival monocyte/macrophage lineage cells sorted from CD45⁺ cells after 5 days of desiccating stress (DS5) ( P adj < 0.001). Genes are organized into five functional categories. The color scale represents z -scores of normalized expression ( blue , low; red , high). These data show desiccating stress–associated shifts in macrophage gene programs and their modulation by NEt-3IB, including enrichment of reparative and growth factor–related genes such as Igf1 . Full gene names for the abbreviations are provided in . ( B ) Representative flow cytometry histograms and quantification validating selected macrophage-associated markers in conjunctival immune cells. ( Left ) Representative histograms of CX3CR1 staining with fluorescence-minus-one (FMO) control, with corresponding quantification of the percentage of CD45⁺CD11b⁺CX3CR1⁺ cells. ( Right ) Representative histograms of IGF-1 staining with FMO control, with corresponding quantification of the percentage of CD45⁺CD11b⁺Mrc1⁺IGF-1⁺ cells. Compared with DS5 no treatment and DS5+vehicle controls, DS5+NEt-3IB increased the proportion ofx CX3CR1⁺ and Mrc1⁺IGF-1⁺ myeloid cells. Each dot represents one biological replicate; bars show mean ± SEM. Statistical significance is indicated as shown: ** P < 0.01; **** P < 0.001; **** P < 0.0001; ns, not significant. ( C ) Representative immunofluorescence images showing IGF-1R/WGA/DAPI staining of wholemount conjunctiva showing surface view ( top ) and βIII-tubulin/IGF-1R/DAPI staining in the cornea ( bottom ). IGF-1R localization is shown because IGF-1 is produced by ocular surface resident macrophages, suggesting a potential macrophage-derived IGF-1/IGF-1R signaling axis acting on ocular surface epithelial and neural compartments during DS. Scale bar : 100 µm.

    Techniques Used: Expressing, Derivative Assay, Functional Assay, Flow Cytometry, Staining, Fluorescence, Control, Immunofluorescence, Produced



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    Effects of rexinoid treatment on macrophage gene and protein expression in the desiccation stress dry eye model and potential macrophage-derived <t>IGF-1/IGF-1R</t> signaling axis. ( A ) Heatmaps showing z -score–scaled expression of selected latent-time–associated genes in four treatment groups (NS, None [untreated], Veh, and NEt-3IB) in conjunctival monocyte/macrophage lineage cells sorted from CD45⁺ cells after 5 days of desiccating stress (DS5) ( P adj < 0.001). Genes are organized into five functional categories. The color scale represents z -scores of normalized expression ( blue , low; red , high). These data show desiccating stress–associated shifts in macrophage gene programs and their modulation by NEt-3IB, including enrichment of reparative and growth factor–related genes such as Igf1 . Full gene names for the abbreviations are provided in . ( B ) Representative flow cytometry histograms and quantification validating selected macrophage-associated markers in conjunctival immune cells. ( Left ) Representative histograms of CX3CR1 staining with fluorescence-minus-one (FMO) control, with corresponding quantification of the percentage of CD45⁺CD11b⁺CX3CR1⁺ cells. ( Right ) Representative histograms of IGF-1 staining with FMO control, with corresponding quantification of the percentage of CD45⁺CD11b⁺Mrc1⁺IGF-1⁺ cells. Compared with DS5 no treatment and DS5+vehicle controls, DS5+NEt-3IB increased the proportion ofx CX3CR1⁺ and Mrc1⁺IGF-1⁺ myeloid cells. Each dot represents one biological replicate; bars show mean ± SEM. Statistical significance is indicated as shown: ** P < 0.01; **** P < 0.001; **** P < 0.0001; ns, not significant. ( C ) Representative immunofluorescence images showing IGF-1R/WGA/DAPI staining of wholemount conjunctiva showing surface view ( top ) and βIII-tubulin/IGF-1R/DAPI staining in the cornea ( bottom ). IGF-1R localization is shown because IGF-1 is produced by ocular surface resident macrophages, suggesting a potential macrophage-derived IGF-1/IGF-1R signaling axis acting on ocular surface epithelial and neural compartments during DS. Scale bar : 100 µm.
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    Effects of rexinoid treatment on macrophage gene and protein expression in the desiccation stress dry eye model and potential macrophage-derived <t>IGF-1/IGF-1R</t> signaling axis. ( A ) Heatmaps showing z -score–scaled expression of selected latent-time–associated genes in four treatment groups (NS, None [untreated], Veh, and NEt-3IB) in conjunctival monocyte/macrophage lineage cells sorted from CD45⁺ cells after 5 days of desiccating stress (DS5) ( P adj < 0.001). Genes are organized into five functional categories. The color scale represents z -scores of normalized expression ( blue , low; red , high). These data show desiccating stress–associated shifts in macrophage gene programs and their modulation by NEt-3IB, including enrichment of reparative and growth factor–related genes such as Igf1 . Full gene names for the abbreviations are provided in . ( B ) Representative flow cytometry histograms and quantification validating selected macrophage-associated markers in conjunctival immune cells. ( Left ) Representative histograms of CX3CR1 staining with fluorescence-minus-one (FMO) control, with corresponding quantification of the percentage of CD45⁺CD11b⁺CX3CR1⁺ cells. ( Right ) Representative histograms of IGF-1 staining with FMO control, with corresponding quantification of the percentage of CD45⁺CD11b⁺Mrc1⁺IGF-1⁺ cells. Compared with DS5 no treatment and DS5+vehicle controls, DS5+NEt-3IB increased the proportion ofx CX3CR1⁺ and Mrc1⁺IGF-1⁺ myeloid cells. Each dot represents one biological replicate; bars show mean ± SEM. Statistical significance is indicated as shown: ** P < 0.01; **** P < 0.001; **** P < 0.0001; ns, not significant. ( C ) Representative immunofluorescence images showing IGF-1R/WGA/DAPI staining of wholemount conjunctiva showing surface view ( top ) and βIII-tubulin/IGF-1R/DAPI staining in the cornea ( bottom ). IGF-1R localization is shown because IGF-1 is produced by ocular surface resident macrophages, suggesting a potential macrophage-derived IGF-1/IGF-1R signaling axis acting on ocular surface epithelial and neural compartments during DS. Scale bar : 100 µm.
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    Insulin-like growth factor-1 <t>(IGF-1R)</t> expression on neurons is elevated after hypoxic-ischemic (HI) challenge. (A) Real-time PCR (RT-PCR) analysis of IGF-1R in neonatal brain at 7 days after HI or sham operation ( n = 6/group). (B) Representative FACS analysis (left) and statistical analysis (right) of IGF-1R + neurons in neonatal brain at 7 days after HI or sham operation ( n = 6/group). (C) Representative images (left) and statistical analysis (right) of Neun (green) and IGF-1R (red) in the cerebral cortex of mice at 7 days after HI or sham operation ( n = 6/group). Scale bar, 25 μm. Data are mean ± SEM. Statistical differences between two groups were evaluated by two-tailed Student’s t -test. * P < 0.05; ** P < 0.01.
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    Image Search Results


    Effects of rexinoid treatment on macrophage gene and protein expression in the desiccation stress dry eye model and potential macrophage-derived IGF-1/IGF-1R signaling axis. ( A ) Heatmaps showing z -score–scaled expression of selected latent-time–associated genes in four treatment groups (NS, None [untreated], Veh, and NEt-3IB) in conjunctival monocyte/macrophage lineage cells sorted from CD45⁺ cells after 5 days of desiccating stress (DS5) ( P adj < 0.001). Genes are organized into five functional categories. The color scale represents z -scores of normalized expression ( blue , low; red , high). These data show desiccating stress–associated shifts in macrophage gene programs and their modulation by NEt-3IB, including enrichment of reparative and growth factor–related genes such as Igf1 . Full gene names for the abbreviations are provided in . ( B ) Representative flow cytometry histograms and quantification validating selected macrophage-associated markers in conjunctival immune cells. ( Left ) Representative histograms of CX3CR1 staining with fluorescence-minus-one (FMO) control, with corresponding quantification of the percentage of CD45⁺CD11b⁺CX3CR1⁺ cells. ( Right ) Representative histograms of IGF-1 staining with FMO control, with corresponding quantification of the percentage of CD45⁺CD11b⁺Mrc1⁺IGF-1⁺ cells. Compared with DS5 no treatment and DS5+vehicle controls, DS5+NEt-3IB increased the proportion ofx CX3CR1⁺ and Mrc1⁺IGF-1⁺ myeloid cells. Each dot represents one biological replicate; bars show mean ± SEM. Statistical significance is indicated as shown: ** P < 0.01; **** P < 0.001; **** P < 0.0001; ns, not significant. ( C ) Representative immunofluorescence images showing IGF-1R/WGA/DAPI staining of wholemount conjunctiva showing surface view ( top ) and βIII-tubulin/IGF-1R/DAPI staining in the cornea ( bottom ). IGF-1R localization is shown because IGF-1 is produced by ocular surface resident macrophages, suggesting a potential macrophage-derived IGF-1/IGF-1R signaling axis acting on ocular surface epithelial and neural compartments during DS. Scale bar : 100 µm.

    Journal: Investigative Ophthalmology & Visual Science

    Article Title: Rexinoid NEt-3IB Promotes Resident Macrophage Gene Expression and Mitigates Desiccation-Induced Ocular Surface Disease

    doi: 10.1167/iovs.67.4.31

    Figure Lengend Snippet: Effects of rexinoid treatment on macrophage gene and protein expression in the desiccation stress dry eye model and potential macrophage-derived IGF-1/IGF-1R signaling axis. ( A ) Heatmaps showing z -score–scaled expression of selected latent-time–associated genes in four treatment groups (NS, None [untreated], Veh, and NEt-3IB) in conjunctival monocyte/macrophage lineage cells sorted from CD45⁺ cells after 5 days of desiccating stress (DS5) ( P adj < 0.001). Genes are organized into five functional categories. The color scale represents z -scores of normalized expression ( blue , low; red , high). These data show desiccating stress–associated shifts in macrophage gene programs and their modulation by NEt-3IB, including enrichment of reparative and growth factor–related genes such as Igf1 . Full gene names for the abbreviations are provided in . ( B ) Representative flow cytometry histograms and quantification validating selected macrophage-associated markers in conjunctival immune cells. ( Left ) Representative histograms of CX3CR1 staining with fluorescence-minus-one (FMO) control, with corresponding quantification of the percentage of CD45⁺CD11b⁺CX3CR1⁺ cells. ( Right ) Representative histograms of IGF-1 staining with FMO control, with corresponding quantification of the percentage of CD45⁺CD11b⁺Mrc1⁺IGF-1⁺ cells. Compared with DS5 no treatment and DS5+vehicle controls, DS5+NEt-3IB increased the proportion ofx CX3CR1⁺ and Mrc1⁺IGF-1⁺ myeloid cells. Each dot represents one biological replicate; bars show mean ± SEM. Statistical significance is indicated as shown: ** P < 0.01; **** P < 0.001; **** P < 0.0001; ns, not significant. ( C ) Representative immunofluorescence images showing IGF-1R/WGA/DAPI staining of wholemount conjunctiva showing surface view ( top ) and βIII-tubulin/IGF-1R/DAPI staining in the cornea ( bottom ). IGF-1R localization is shown because IGF-1 is produced by ocular surface resident macrophages, suggesting a potential macrophage-derived IGF-1/IGF-1R signaling axis acting on ocular surface epithelial and neural compartments during DS. Scale bar : 100 µm.

    Article Snippet: The following primary antibodies were used: βIII-tubulin (ab215037; Abcam, Cambridge, UK) and human/mouse IGF-1R (AF-305-NA; R&D Systems, Minneapolis, MN, USA).

    Techniques: Expressing, Derivative Assay, Functional Assay, Flow Cytometry, Staining, Fluorescence, Control, Immunofluorescence, Produced

    Representative images of IHC staining. IHC staining of TNBC specimens was performed as described in the Methods section with antibodies to IR, IGF-1R, pErk1/2, and FOXO3a. The images are shown in 40× magnification. Abbreviations: IGF-1R, IGR-1 receptor; IHC, immunohistochemistry; IR, insulin receptor; pErk1/2, phosphorylated Erk1/2; TNBC, triple negative breast cancer.

    Journal: Journal of the Endocrine Society

    Article Title: Insulin receptor expression and its association with hyperinsulinemia in triple negative breast cancer

    doi: 10.1210/jendso/bvag041

    Figure Lengend Snippet: Representative images of IHC staining. IHC staining of TNBC specimens was performed as described in the Methods section with antibodies to IR, IGF-1R, pErk1/2, and FOXO3a. The images are shown in 40× magnification. Abbreviations: IGF-1R, IGR-1 receptor; IHC, immunohistochemistry; IR, insulin receptor; pErk1/2, phosphorylated Erk1/2; TNBC, triple negative breast cancer.

    Article Snippet: Primary antibodies were anti-IR Ab (ab137747, Abcam, Cambridge, MA, RRID:AB_3717506) 1:300 dilution, anti-IGF-1R Ab (#3027, Cell Signaling, Danvers, CA, RRID:AB_2122378) 1:200 dilution, anti-FOXO3a Ab (#12829, Cell Signaling, RRID:AB_2636990) 1:3200 dilution, and anti-pErk1/2 (Thr202/Tyr204) Ab (#4370, Cell Signaling, RRID:AB_2315112) 1:400 dilution.

    Techniques: Immunohistochemistry

    Representative images of protein localization for IR, IGF1R, FOXO3a, and pErk1/2. IHC staining of TNBC specimens was performed and evaluated for localization of proteins. The images are shown in 40× magnification. Abbreviations: IGF-1R, IGR-1 receptor; IHC, immunohistochemistry; IR, insulin receptor; pErk1/2, phosphorylated Erk1/2; TNBC, triple negative breast cancer.

    Journal: Journal of the Endocrine Society

    Article Title: Insulin receptor expression and its association with hyperinsulinemia in triple negative breast cancer

    doi: 10.1210/jendso/bvag041

    Figure Lengend Snippet: Representative images of protein localization for IR, IGF1R, FOXO3a, and pErk1/2. IHC staining of TNBC specimens was performed and evaluated for localization of proteins. The images are shown in 40× magnification. Abbreviations: IGF-1R, IGR-1 receptor; IHC, immunohistochemistry; IR, insulin receptor; pErk1/2, phosphorylated Erk1/2; TNBC, triple negative breast cancer.

    Article Snippet: Primary antibodies were anti-IR Ab (ab137747, Abcam, Cambridge, MA, RRID:AB_3717506) 1:300 dilution, anti-IGF-1R Ab (#3027, Cell Signaling, Danvers, CA, RRID:AB_2122378) 1:200 dilution, anti-FOXO3a Ab (#12829, Cell Signaling, RRID:AB_2636990) 1:3200 dilution, and anti-pErk1/2 (Thr202/Tyr204) Ab (#4370, Cell Signaling, RRID:AB_2315112) 1:400 dilution.

    Techniques: Immunohistochemistry

    Overall survival for women with breast cancer based on IR and IGF-1R protein expression in KMPlot.com . KMPlot.com was used to generate overall survival Kaplan-Meier plots. The dataset used was not from our cohort but from Tang and colleagues ; n = 65 patients had IR (A) and IGF1R (B) protein expression available. The patients were split using the “auto select best cutoff” percentile option in KMPlot. No other restrictions were added to the analysis. The plots show hazard ratios with 95% confidence intervals in parentheses . Abbreviations: IGF-1R, IGR-1 receptor; IR, insulin receptor.

    Journal: Journal of the Endocrine Society

    Article Title: Insulin receptor expression and its association with hyperinsulinemia in triple negative breast cancer

    doi: 10.1210/jendso/bvag041

    Figure Lengend Snippet: Overall survival for women with breast cancer based on IR and IGF-1R protein expression in KMPlot.com . KMPlot.com was used to generate overall survival Kaplan-Meier plots. The dataset used was not from our cohort but from Tang and colleagues ; n = 65 patients had IR (A) and IGF1R (B) protein expression available. The patients were split using the “auto select best cutoff” percentile option in KMPlot. No other restrictions were added to the analysis. The plots show hazard ratios with 95% confidence intervals in parentheses . Abbreviations: IGF-1R, IGR-1 receptor; IR, insulin receptor.

    Article Snippet: Primary antibodies were anti-IR Ab (ab137747, Abcam, Cambridge, MA, RRID:AB_3717506) 1:300 dilution, anti-IGF-1R Ab (#3027, Cell Signaling, Danvers, CA, RRID:AB_2122378) 1:200 dilution, anti-FOXO3a Ab (#12829, Cell Signaling, RRID:AB_2636990) 1:3200 dilution, and anti-pErk1/2 (Thr202/Tyr204) Ab (#4370, Cell Signaling, RRID:AB_2315112) 1:400 dilution.

    Techniques: Expressing

    MSC therapy enhances growth factor expression and receptor signaling in SI-injured R28 cells and TBI rats. (A) Representative growth factor antibody array images from conditioned media of R28 cells subjected to stretch injury (SI), with or without MSC coculture. Increased secretion of HGF, IGFBP-4, IGFBP-6, and VEGF was observed in the SI+MSC group. Positive controls are marked in red boxes; significantly regulated proteins are highlighted in blue. Quantitative analysis of mean pixel density is shown. (B) ELISA quantification of serum VEGF, HGF, IGFBP-4, and IGFBP-6 levels in TBI rats. TBI reduced systemic levels of these growth factors, while MSC treatment significantly restored them (except IGFBP-6). (C) ELISA quantification of VEGF, HGF, IGFBP-4, and IGFBP-6 in whole eyeball lysates (OD and OS). Due to the technical difficulty in isolating the retina alone, entire ocular globes were homogenized for assay. TBI significantly reduced VEGF, HGF, and IGFBP-6 levels in the eye, while MSC therapy markedly reversed these changes (except VEGF). (D) Western blot analysis of IGF-1R, VEGFR2, phosphorylated VEGFR2 (p-VEGFR2), and c-Met in R28 cell lysates. SI reduced receptor expression, while MSC coculture significantly restored it. Densitometric quantification is normalized to β-actin. (E) Western blot analysis of downstream signaling proteins in R28 cells. MSC coculture upregulated phosphorylated Akt (p-Akt), Sirtuin 1 (SirT1), and β-catenin in SI-injured cells. Quantified values are normalized to β-actin (p < 0.0001). Abbreviations: HGF, hepatocyte growth factor; IGFBP-4/6, insulin-like growth factor binding protein-4/6; VEGF, vascular endothelial growth factor; IGF-1R, insulin-like growth factor 1 receptor; VEGFR2, vascular endothelial growth factor receptor 2; p-Akt, phosphorylated Akt; SirT1, Sirtuin 1. Data are presented as mean ± SD. In vitro experiments (A, D, E) were performed in at least three independent replicates. In vivo data (B, C) are from n=8-9 (serum ELISA) or n = 6 (ocular tissues ELISA) animals per group. For original blot images, see the Supplementary Data file.

    Journal: International Journal of Medical Sciences

    Article Title: Stem cells ameliorate neurotrauma-induced visual disturbances and retinal degeneration via broad normalization of β-catenin-related signaling

    doi: 10.7150/ijms.123975

    Figure Lengend Snippet: MSC therapy enhances growth factor expression and receptor signaling in SI-injured R28 cells and TBI rats. (A) Representative growth factor antibody array images from conditioned media of R28 cells subjected to stretch injury (SI), with or without MSC coculture. Increased secretion of HGF, IGFBP-4, IGFBP-6, and VEGF was observed in the SI+MSC group. Positive controls are marked in red boxes; significantly regulated proteins are highlighted in blue. Quantitative analysis of mean pixel density is shown. (B) ELISA quantification of serum VEGF, HGF, IGFBP-4, and IGFBP-6 levels in TBI rats. TBI reduced systemic levels of these growth factors, while MSC treatment significantly restored them (except IGFBP-6). (C) ELISA quantification of VEGF, HGF, IGFBP-4, and IGFBP-6 in whole eyeball lysates (OD and OS). Due to the technical difficulty in isolating the retina alone, entire ocular globes were homogenized for assay. TBI significantly reduced VEGF, HGF, and IGFBP-6 levels in the eye, while MSC therapy markedly reversed these changes (except VEGF). (D) Western blot analysis of IGF-1R, VEGFR2, phosphorylated VEGFR2 (p-VEGFR2), and c-Met in R28 cell lysates. SI reduced receptor expression, while MSC coculture significantly restored it. Densitometric quantification is normalized to β-actin. (E) Western blot analysis of downstream signaling proteins in R28 cells. MSC coculture upregulated phosphorylated Akt (p-Akt), Sirtuin 1 (SirT1), and β-catenin in SI-injured cells. Quantified values are normalized to β-actin (p < 0.0001). Abbreviations: HGF, hepatocyte growth factor; IGFBP-4/6, insulin-like growth factor binding protein-4/6; VEGF, vascular endothelial growth factor; IGF-1R, insulin-like growth factor 1 receptor; VEGFR2, vascular endothelial growth factor receptor 2; p-Akt, phosphorylated Akt; SirT1, Sirtuin 1. Data are presented as mean ± SD. In vitro experiments (A, D, E) were performed in at least three independent replicates. In vivo data (B, C) are from n=8-9 (serum ELISA) or n = 6 (ocular tissues ELISA) animals per group. For original blot images, see the Supplementary Data file.

    Article Snippet: IGF-1R , 1:2000 , Cell Signaling Technology , 3027 , WB.

    Techniques: Expressing, Ab Array, Enzyme-linked Immunosorbent Assay, Western Blot, Binding Assay, In Vitro, In Vivo

    MSC therapy restores TBI-induced neurotrophic growth factor signaling downregulation in retinal neurons. (A) Representative immunofluorescence images of retinal sections (400× magnification) from the indicated groups (Sham+Veh, Sham+MSC, TBI+Veh, TBI+MSC). Images show neuronal co-expression of growth factor ligands and receptors. Top panel: Triple staining for NeuN (neuronal marker, gray), c-Met (HGF receptor, green), and HGF (red). Second panel: NeuN, IGF-1R (IGF receptor, green), and IGFBP-4 (red). Third panel: NeuN, IGF-1R (green), and IGFBP-6 (red). Bottom panel: NeuN, VEGFR2 (VEGF receptor, green), and VEGF (red). DAPI (blue) stains nuclei. Retinal regions include ipsilesional OD-T, OS-N and contralesional OD-N, OS-T segments. (B-E) Quantification of triple-positive cells (NeuN+ growth factor receptor+ ligand) per retinal section: (B) NeuN+c-Met+HGF, (C) NeuN+IGF-1R+IGFBP-4, (D) NeuN+IGF-1R+IGFBP-6, (E) NeuN+VEGFR2+VEGFA. Data were obtained from 6 animals in each group and expressed as mean±SD.

    Journal: International Journal of Medical Sciences

    Article Title: Stem cells ameliorate neurotrauma-induced visual disturbances and retinal degeneration via broad normalization of β-catenin-related signaling

    doi: 10.7150/ijms.123975

    Figure Lengend Snippet: MSC therapy restores TBI-induced neurotrophic growth factor signaling downregulation in retinal neurons. (A) Representative immunofluorescence images of retinal sections (400× magnification) from the indicated groups (Sham+Veh, Sham+MSC, TBI+Veh, TBI+MSC). Images show neuronal co-expression of growth factor ligands and receptors. Top panel: Triple staining for NeuN (neuronal marker, gray), c-Met (HGF receptor, green), and HGF (red). Second panel: NeuN, IGF-1R (IGF receptor, green), and IGFBP-4 (red). Third panel: NeuN, IGF-1R (green), and IGFBP-6 (red). Bottom panel: NeuN, VEGFR2 (VEGF receptor, green), and VEGF (red). DAPI (blue) stains nuclei. Retinal regions include ipsilesional OD-T, OS-N and contralesional OD-N, OS-T segments. (B-E) Quantification of triple-positive cells (NeuN+ growth factor receptor+ ligand) per retinal section: (B) NeuN+c-Met+HGF, (C) NeuN+IGF-1R+IGFBP-4, (D) NeuN+IGF-1R+IGFBP-6, (E) NeuN+VEGFR2+VEGFA. Data were obtained from 6 animals in each group and expressed as mean±SD.

    Article Snippet: IGF-1R , 1:2000 , Cell Signaling Technology , 3027 , WB.

    Techniques: Immunofluorescence, Expressing, Staining, Marker

    Insulin-like growth factor-1 (IGF-1R) expression on neurons is elevated after hypoxic-ischemic (HI) challenge. (A) Real-time PCR (RT-PCR) analysis of IGF-1R in neonatal brain at 7 days after HI or sham operation ( n = 6/group). (B) Representative FACS analysis (left) and statistical analysis (right) of IGF-1R + neurons in neonatal brain at 7 days after HI or sham operation ( n = 6/group). (C) Representative images (left) and statistical analysis (right) of Neun (green) and IGF-1R (red) in the cerebral cortex of mice at 7 days after HI or sham operation ( n = 6/group). Scale bar, 25 μm. Data are mean ± SEM. Statistical differences between two groups were evaluated by two-tailed Student’s t -test. * P < 0.05; ** P < 0.01.

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Neuroprotection of IGF-1 in neonatal hypoxic-ischemic brain injury through downregulation of FoXO3a-PUMA pathway

    doi: 10.3389/fncel.2025.1685800

    Figure Lengend Snippet: Insulin-like growth factor-1 (IGF-1R) expression on neurons is elevated after hypoxic-ischemic (HI) challenge. (A) Real-time PCR (RT-PCR) analysis of IGF-1R in neonatal brain at 7 days after HI or sham operation ( n = 6/group). (B) Representative FACS analysis (left) and statistical analysis (right) of IGF-1R + neurons in neonatal brain at 7 days after HI or sham operation ( n = 6/group). (C) Representative images (left) and statistical analysis (right) of Neun (green) and IGF-1R (red) in the cerebral cortex of mice at 7 days after HI or sham operation ( n = 6/group). Scale bar, 25 μm. Data are mean ± SEM. Statistical differences between two groups were evaluated by two-tailed Student’s t -test. * P < 0.05; ** P < 0.01.

    Article Snippet: To examine the percentages of IGF-1 + or IGF-1R + cells, cells were stained with a primary anti-IGF-1 (SinoBiological) or anti-IGF-1R antibody (R&D Systems) followed by PE- or FITC-conjugated secondary antibodies (eBioscience).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Two Tailed Test