Journal: Journal of Cellular and Molecular Medicine

Article Title: Lycopene inhibits endothelial‐to‐mesenchymal transition of choroidal vascular endothelial cells in laser‐induced mouse choroidal neovascularization

doi: 10.1111/jcmm.17730

Figure Lengend Snippet: The information of PCR primers used in the study was shown

Article Snippet: Human PEDF recombinant protein (1177‐SF, R&D Systems, USA) was added to HCVECs at the concentration of 2 μg/mL for 48 h.

Techniques: Sequencing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Lycopene inhibits endothelial‐to‐mesenchymal transition of choroidal vascular endothelial cells in laser‐induced mouse choroidal neovascularization

doi: 10.1111/jcmm.17730

Figure Lengend Snippet: LYC‐down‐regulated AR and ‐induced MITF up‐regulate PEDF transcription and expression in HCVECs under hypoxic conditions. HCVECs have been classified into normal, hypoxia, hypoxia +0.1% DMSO, hypoxia + LYC (20 μM), hypoxia + LYC + TES, and hypoxia + LYC + ML329 groups. (A) PEDF mRNA levels were detected by qRT‐PCR. (B) PEDF protein levels were detected by WB. (C) PEDF protein levels were analysed. (D) The MITF binding site (5′‐CACGTG‐3′, from −672 to −667) in the human PEDF promoter is shown. (E) The activity of the human PEDF promoter was detected by luciferase reporter assay in HCVECs. (F) The direct binding of MITF to the PEDF promoter was detected by CHIP. * p < 0.05, compared to the normal group. # p < 0.05, in comparison to the hypoxia group. % p < 0.05, in comparison to the hypoxia + LYC group. n = 5 in each group.

Article Snippet: Human PEDF recombinant protein (1177‐SF, R&D Systems, USA) was added to HCVECs at the concentration of 2 μg/mL for 48 h.

Techniques: Expressing, Quantitative RT-PCR, Binding Assay, Activity Assay, Luciferase, Reporter Assay, Comparison

Journal: Journal of Cellular and Molecular Medicine

Article Title: Lycopene inhibits endothelial‐to‐mesenchymal transition of choroidal vascular endothelial cells in laser‐induced mouse choroidal neovascularization

doi: 10.1111/jcmm.17730

Figure Lengend Snippet: LYC‐induced PEDF binds to LR on HCVECs under hypoxic conditions. HCVECs were divided into normal, hypoxia, hypoxia +0.1% DMSO, hypoxia + LYC (20 μM), hypoxia + LYC + control siRNA, hypoxia + LYC + PEDF siRNA and hypoxia + human PEDF recombinant protein groups. (A) PEDF‐R and LR protein levels were detected by WB. (B‐C) PEDF‐R and LR protein levels were analysed. (D) HCVECs were divided into normal, hypoxia, hypoxia +0.1% DMSO and hypoxia + LYC groups. PEDF and LR were labelled in CECs. (E) The co‐localization of PEDF and LR was analysed. * p < 0.05, in comparison to the normal group. # p < 0.05, in comparison to the hypoxia group. % p < 0.05, in comparison to the hypoxia + LYC group. n.s. denoted no significance. n = 5 in each group.

Article Snippet: Human PEDF recombinant protein (1177‐SF, R&D Systems, USA) was added to HCVECs at the concentration of 2 μg/mL for 48 h.

Techniques: Control, Recombinant, Comparison

Journal: Journal of Cellular and Molecular Medicine

Article Title: Lycopene inhibits endothelial‐to‐mesenchymal transition of choroidal vascular endothelial cells in laser‐induced mouse choroidal neovascularization

doi: 10.1111/jcmm.17730

Figure Lengend Snippet: LYC‐induced PEDF inhibits EndMT of HCVECs via down‐regulating AKT/β‐catenin pathway under hypoxic conditions. HCVECs were divided into normal, hypoxia, hypoxia +0.1% DMSO, hypoxia + LYC (20 μM), hypoxia + LYC + control siRNA, hypoxia + LYC + PEDF siRNA, hypoxia + LYC + SC79, and hypoxia + LYC + SKL2001 groups. (A) P‐AKT, AKT, p‐β‐catenin and β‐catenin protein levels were detected by WB. (B, C) The relative expression of p‐AKT and p‐β‐catenin was analysed. (D) CD31, VE‐cadherin, α‐SMA and vimentin protein levels were detected by WB. (E–H) The relative expression of each molecule was analysed. * p < 0.05, in comparison to the normal group. # p < 0.05, in comparison to the hypoxia group. % p < 0.05, in comparison to the hypoxia + LYC group, n = 5 in each group.

Article Snippet: Human PEDF recombinant protein (1177‐SF, R&D Systems, USA) was added to HCVECs at the concentration of 2 μg/mL for 48 h.

Techniques: Control, Expressing, Comparison

Journal: Journal of Cellular and Molecular Medicine

Article Title: Lycopene inhibits endothelial‐to‐mesenchymal transition of choroidal vascular endothelial cells in laser‐induced mouse choroidal neovascularization

doi: 10.1111/jcmm.17730

Figure Lengend Snippet: LYC alleviates mouse laser‐induced subretinal fibrosis secondary to CNV via up‐regulating PEDF. The mice were divided into normal, CNV 14 days, CNV 14 days + 0.1% DMSO (oral gavage; 0.8 mg/kg/day for consecutive 14 days), CNV 14 days + LYC (oral gavage; 0.8 mg/kg/day for consecutive 14 days), CNV 14 days + LYC + control siRNA, CNV 14 days + LYC + PEDF siRNA, and CNV 14 days + CON groups. (A) Representative images of subretinal fibrosis lesions by marking α‐SMA in each group (except the normal group) at post‐laser 14 days were shown. Scale bar = 100 μm. (B) Quantitative analyses for the area of α‐SMA were done. Snail (C) and α‐SMA (D) mRNA levels were detected by qRT‐PCR. (E) Snail and α‐SMA protein levels were detected by Western blot. Snail (F) and α‐SMA (G) protein levels were analysed. * p < 0.05, in comparison to the normal group. # p < 0.05, in comparison to the hypoxia group. % p < 0.05, in comparison to the hypoxia + LYC group. n = 5 in each group.

Article Snippet: Human PEDF recombinant protein (1177‐SF, R&D Systems, USA) was added to HCVECs at the concentration of 2 μg/mL for 48 h.

Techniques: Control, Quantitative RT-PCR, Western Blot, Comparison

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