pedf  (R&D Systems)

Journal: Journal of nanobiotechnology

Article Title: Enhanced therapeutic effect of PEDF-loaded mesenchymal stem cell-derived small extracellular vesicles against oxygen-induced retinopathy through increased stability and penetrability of PEDF.

doi: 10.1186/s12951-023-02066-z

Figure Lengend Snippet: Fig. 1 Characterisation of MSCs and sEVs. A Representative images of adipogenic (oil red O staining), osteogenic (alizarin red S staining), and chondrogenic (alcian blue staining) differentiation assay. B Analysis of sizes of sEVs from each group using Nanosight. C Representative transmission electron micrograph for each group; scale bar = 200 nm. D Representative western blots for loaded proteins and markers (PEDF, CD63, TSG101, and CD9) in MSC-sEVs. E Flow cytometry for measuring the loading efficiency of PEDF in PEDF-sEVs. FITC-labelled PEDF was used. F The concentrations of PEDF in sEVs and PEDF-sEVs were determined using ELISA (n = 3/group). The data are represented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001

Article Snippet: PEDF (1177- SF-025, R&D SYSTEMS, USA) was mixed with sEVs at a 1:5 concentration ratio.

Techniques: Staining, Differentiation Assay, Transmission Assay, Western Blot, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Journal of nanobiotechnology

Article Title: Enhanced therapeutic effect of PEDF-loaded mesenchymal stem cell-derived small extracellular vesicles against oxygen-induced retinopathy through increased stability and penetrability of PEDF.

doi: 10.1186/s12951-023-02066-z

Figure Lengend Snippet: Fig. 3 PEDF-sEVs suppress the expression of inflammatory cytokines in HRECs. A Starved HRECs were pre-treated with PEDF, sEVs, or PEDF-sEVs under stimulation with 10 ng/mL VEGF for 24 h. Representative western blot images showing the expression of ICAM-1 in HRECs. B Relative expression of ICAM-1 compared with that of GAPDH (n = 3). C, D, E. PCR analysis of the expression of TNF-α, VCAM-1, and IL-1β in HRECs (n = 5). The data are represented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001

Article Snippet: PEDF (1177- SF-025, R&D SYSTEMS, USA) was mixed with sEVs at a 1:5 concentration ratio.

Techniques: Expressing, Western Blot

Journal: Journal of clinical & experimental ophthalmology

Article Title: Pressure-Induced Alterations in PEDF and PEDF-R Expression: Implications for Neuroprotective Signaling in Glaucoma

doi: 10.4172/2155-9570.1000491

Figure Lengend Snippet: Primary antibodies used for immunohistochemistry and immunocytochemistry. Primary antibodies, sources, and concentrations used for detection of PEDF, PEDF-R and labeling of RGCs and Müller cells in murine retina.

Article Snippet: Sections were incubated overnight at 4°C in primary antibody solution containing anti-PEDF antibody ( ) with Recombinant PEDF protein (20 μg/ml; R&D Systems, catalog #1177-SF-025).

Techniques: Immunohistochemistry, Immunocytochemistry, Labeling, Concentration Assay

Journal: Journal of clinical & experimental ophthalmology

Article Title: Pressure-Induced Alterations in PEDF and PEDF-R Expression: Implications for Neuroprotective Signaling in Glaucoma

doi: 10.4172/2155-9570.1000491

Figure Lengend Snippet: PEDF is constitutively expressed in healthy retina. A. Graphical representation of pedf total mRNA levels in naïve C57 retina as measured by qRT-PCR. Y-axis represents the average delta of threshold cycle (Δ-CT) values for pedf normalized to the control gene gapdh. B . Representative micrograph of longitudinal sections of retina from naïve mice immunolabeled with antibody against PEDF with (right panel) and without (left) pre-absorption with recombinant PEDF (rPEDF). Morphology consistent with localization to RGCs (white arrowheads) and Müller cells (black arrowheads). C . Retinal layer-specific quantification of PEDF labeling, expressed in intensity (arbitrary units) per area (mm 2 ). All asterisks denote p<0.05. ONL: Outer Nuclear Layer; OPL: Outer Plexiform Layer; INL: Inner Nuclear Layer; IPL: Inner Plexiform Layer; GCL: Ganglion Cell Layer; NFL: Nerve Fiber Layer.

Article Snippet: Sections were incubated overnight at 4°C in primary antibody solution containing anti-PEDF antibody ( ) with Recombinant PEDF protein (20 μg/ml; R&D Systems, catalog #1177-SF-025).

Techniques: Quantitative RT-PCR, Control, Immunolabeling, Recombinant, Labeling

Journal: Journal of clinical & experimental ophthalmology

Article Title: Pressure-Induced Alterations in PEDF and PEDF-R Expression: Implications for Neuroprotective Signaling in Glaucoma

doi: 10.4172/2155-9570.1000491

Figure Lengend Snippet: PEDF is associated with RGCs and Müller cell endfeet in the ganglion cell and nerve fiber layers. A . Representative confocal micrographs with orthogonal view (bottom panel) of wholemount retina from naïve mice co-immunolabeled with antibodies against PEDF (green) and the RGC marker γ-synuclein (red). PEDF localizes to the area surrounding PEDF γ-synuclein+ RGCs in the ganglion cell layer (GCL; arrowheads). B . Representative confocal micrographs with orthogonal view (bottom panel) of wholemount retina from naïve mice co-immunolabeled with antibodies against PEDF (green) and the Müller cell marker glutamine synthetase (GluSyn; blue). PEDF localizes to the area surrounding Müller cell endfeet in the nerve fiber layer (NFL).

Article Snippet: Sections were incubated overnight at 4°C in primary antibody solution containing anti-PEDF antibody ( ) with Recombinant PEDF protein (20 μg/ml; R&D Systems, catalog #1177-SF-025).

Techniques: Immunolabeling, Marker

Journal: Journal of clinical & experimental ophthalmology

Article Title: Pressure-Induced Alterations in PEDF and PEDF-R Expression: Implications for Neuroprotective Signaling in Glaucoma

doi: 10.4172/2155-9570.1000491

Figure Lengend Snippet: Elevated IOP increases PEDF expression in murine retina. A . Graphical representation of changes in pedf total mRNA in retina from saline- and microbead-injected mice, as measured by qPCR. Y-axis represents the fold-change in Δ-CT values for pedf (normalized to the control gene gapdh) microbead-injected, as compared to saline-injected. B . Representative micrographs of immunolabeling for PEDF in longitudinal sections of retina from saline- (right panel) and microbead-injected (left panel) mice demonstrate increased intensity of labeling in all layers of retina from microbead-injected eyes, as compared to saline-injected eyes. The pattern of PEDF immunolabeling is consistent with RGC soma in the GCL (white arrowheads) and Müller cell processes and endfeet in the ONL and NFL (black arrowheads), respectively. C . Retinal layer-specific quantification of PEDF labeling, expressed in intensity (arbitrary units) per area (mm2). All asterisks denote p<0.05. ONL: Outer Nuclear Layer; OPL: Outer Plexiform Layer; INL: Inner Nuclear Layer; IPL: Inner Plexiform Layer; GCL: Ganglion Cell Layer; NFL: Nerve Fiber Layer

Article Snippet: Sections were incubated overnight at 4°C in primary antibody solution containing anti-PEDF antibody ( ) with Recombinant PEDF protein (20 μg/ml; R&D Systems, catalog #1177-SF-025).

Techniques: Expressing, Saline, Injection, Control, Immunolabeling, Labeling

Journal: Journal of clinical & experimental ophthalmology

Article Title: Pressure-Induced Alterations in PEDF and PEDF-R Expression: Implications for Neuroprotective Signaling in Glaucoma

doi: 10.4172/2155-9570.1000491

Figure Lengend Snippet: Elevated pressure alters PEDF production and localization in RGCs and Müller cells in vitro . A . Graphical representation of PEDF concentration (pg/ml; y-axis) in media from primary cultures of purified RGCs exposed to ambient or elevated pressure for 48 hours, as measured by multiplex ELISA. Asterisk denotes p<0.05. B . Fluorescent micrographs of PEDF (green) and γ-synuclein (red) immunolabeling in purified, primary cultures of RGCs exposed to either ambient (top panels) or elevated (bottom panels) pressure for 48 hours reveals a reduction in localization of PEDF to neurites following exposure to elevated pressure (black arrowheads) versus ambient pressure (white arrowheads). C . Graphical representation of PEDF concentration (pg/ml; y-axis) in media from primary cultures of purified Müller cells exposed to ambient or elevated pressure for 48 hours, as measured by multiplex ELISA. D . Fluorescent micrographs of PEDF (green) and glutamine synthetase (red) immunolabeling in purified, primary cultures of Müller cells exposed to either ambient (top panels) or elevated (bottom panels) pressure for 48 hours reveals retraction in cellular processes that is associated with a reduction in PEDF staining following exposure to elevated pressure (black arrowheads), as compared to ambient pressure (white arrowheads).

Article Snippet: Sections were incubated overnight at 4°C in primary antibody solution containing anti-PEDF antibody ( ) with Recombinant PEDF protein (20 μg/ml; R&D Systems, catalog #1177-SF-025).

Techniques: In Vitro, Concentration Assay, Purification, Multiplex Assay, Enzyme-linked Immunosorbent Assay, Immunolabeling, Staining

Journal: Journal of clinical & experimental ophthalmology

Article Title: Pressure-Induced Alterations in PEDF and PEDF-R Expression: Implications for Neuroprotective Signaling in Glaucoma

doi: 10.4172/2155-9570.1000491

Figure Lengend Snippet: PEDF-R is constitutively expressed by RGCs and Müller cells in healthy retina. A . Graphical representation of pedf-r total mRNA levels in naïve C57 retina as measured by qRT-PCR. Y-axis represents the average delta of threshold cycle (Δ-CT) values for pedf-r normalized to the control gene gapdh. B . Representative micrograph of longitudinal sections of retina from naïve mice immunolabeled with antibody against PEDF-R. Morphology consistent with localization to RGCs (white arrowheads) and Müller cells (black arrowheads). C . Retinal layer-specific quantification of PEDF-R labeling, expressed in intensity (arbitrary units) per area (mm2). All asterisks denote p<0.05. D . Representative confocal micrographs with orthogonal view (bottom panel) of wholemount retina from naïve mice co-immunolabeled with antibodies against PEDF-R (green) and the RGC marker SMI-31 (red). PEDF-R localizes to SMI-31+RGC soma and axons, as indicated by the yellow appearance of punctate immunolabeling (arrowheads). E . Representative confocal micrographs with orthogonal view (bottom panel) of wholemount retina from naïve mice co-immunolabeled with antibodies against PEDF-R (green) and the Müller cell marker glutamine synthetase (GluSyn; blue). PEDF-R co-localizes with Müller cell endfeet in the NFL, as indicated by the white appearance of immunolabeling (arrowheads). ONL: Outer Nuclear Layer; OPL: Outer Plexiform Layer; INL: Inner Nuclear Layer; IPL: Inner Plexiform Layer; GCL: Ganglion Cell Layer; NFL: Nerve Fiber Layer.

Article Snippet: Sections were incubated overnight at 4°C in primary antibody solution containing anti-PEDF antibody ( ) with Recombinant PEDF protein (20 μg/ml; R&D Systems, catalog #1177-SF-025).

Techniques: Quantitative RT-PCR, Control, Immunolabeling, Labeling, Marker

Journal: Journal of clinical & experimental ophthalmology

Article Title: Pressure-Induced Alterations in PEDF and PEDF-R Expression: Implications for Neuroprotective Signaling in Glaucoma

doi: 10.4172/2155-9570.1000491

Figure Lengend Snippet: Elevated IOP increases PEDF-R expression in murine retina. A . Graphical representation of pedf-r total mRNA in retina from saline- and microbead-injected mice, as measured by qPCR. y-axis represents fold change in Δ-CT values for pedf-r (normalized to the control gene gapdh) for microbead-injected versus saline-injected. Asterisk denotes p<0.05. B . Representative micrographs of immunolabeling for PEDF-R in longitudinal sections of retina from saline- (right panel) and microbead-injected (left panel) mice demonstrate increased intensity of labeling, particularly in the IPL, GCL and NFL, as compared to saline-injected eyes. The pattern of PEDF-R immunolabeling is consistent with RGC soma in the GCL (white arrowheads) and Müller cell processes and endfeet in the ONL and NFL (black arrowheads), respectively. C . Retinal layer-specific quantification of PEDF-R labeling, expressed in intensity (arbitrary units) per area (mm 2 ). All asterisks denote p<0.05. ONL: Outer Nuclear Layer; OPL: Outer Plexiform Layer; INL: Inner Nuclear Layer; IPL: Inner Plexiform Layer; GCL: Ganglion Cell Layer; NFL: Nerve Fiber Layer

Article Snippet: Sections were incubated overnight at 4°C in primary antibody solution containing anti-PEDF antibody ( ) with Recombinant PEDF protein (20 μg/ml; R&D Systems, catalog #1177-SF-025).

Techniques: Expressing, Saline, Injection, Control, Immunolabeling, Labeling

Journal: Journal of clinical & experimental ophthalmology

Article Title: Pressure-Induced Alterations in PEDF and PEDF-R Expression: Implications for Neuroprotective Signaling in Glaucoma

doi: 10.4172/2155-9570.1000491

Figure Lengend Snippet: Elevated pressure alters PEDF-R localization in RGCs and Müller cells in vitro. A . Fluorescent micrographs of PEDF-R (green) and SMI-31 (red) immunolabeling in purified, primary cultures of RGCs exposed to either ambient (top panels) or elevated (bottom panels) pressure for 48 hours reveals a reduction in localization of PEDF-R to neurites (black arrowheads) following exposure to elevated pressure, as compared to ambient pressure (white arrowheads). B . Fluorescent micrographs of PEDF-R (green) and glutamine synthetase (red) immunolabeling in purified, primary cultures of Müller cells exposed to either ambient (top panels) or elevated (bottom panels) pressure for 48 hours reveals a reduction in localization of PEDF-R to processes that is associated with process retraction (black arrowheads) following exposure to elevated pressure (black arrowheads) versus ambient pressure (white arrowheads).

Article Snippet: Sections were incubated overnight at 4°C in primary antibody solution containing anti-PEDF antibody ( ) with Recombinant PEDF protein (20 μg/ml; R&D Systems, catalog #1177-SF-025).

Techniques: In Vitro, Immunolabeling, Purification

Journal: Journal of clinical & experimental ophthalmology

Article Title: Pressure-Induced Alterations in PEDF and PEDF-R Expression: Implications for Neuroprotective Signaling in Glaucoma

doi: 10.4172/2155-9570.1000491

Figure Lengend Snippet: PEDF signaling protects RGCs from pressure-induced apoptosis. A . Graphical representation of the percentage TUNEL+RGCs treated with vehicle or the PEDF-R specific inhibitor aglistatin (50 µM, 100 µM, 200 µM, 400 µM) and exposed to ambient or elevated pressure for 48 hours. Asterisks denote statistical significance (p<0.05) between vehicle and drug treatments within each pressure condition. B . Graphical representation of the same data in (A) depicted as the percentage change in TUNEL+RGCs between ambient and elevated pressure within each drug treatment. Asterisks denote statistical significance between ambient and elevated pressure within each drug treatment (p<0.05). C . Graphical representation of the percentage of TUNEL+RGCs treated with vehicle, 50ng/ml recombinant PEDF (rPEDF) or 100 ng/ml rPEDF and exposed to ambient or elevated pressure for 48 hours. Asterisks denote statistical significance (p<0.05).

Article Snippet: Sections were incubated overnight at 4°C in primary antibody solution containing anti-PEDF antibody ( ) with Recombinant PEDF protein (20 μg/ml; R&D Systems, catalog #1177-SF-025).

Techniques: TUNEL Assay, Recombinant