Journal: Journal of Ginseng Research

Article Title: Ginsenoside Re regulates PFKFB3-mediated glycolysis to inhibit endothelial cell migration to ameliorate atherosclerosis

doi: 10.1016/j.jgr.2025.11.012

Figure Lengend Snippet: Re suppresses ox-LDL-induced HUVEC proliferation and migration. (A, B) Results of the CCK-8 assay, n = 6. (C) Representative images of HUVECs at 0 h and 12 h after ox-LDL induction in wound healing experiments, bar = 50 μm. (D) Quantification of EC migration in the wound healing assay, n = 3. (E) Western blot assay and quantitative data of VE-cadherin and VEGFR2 in HUVECs, n = 3. ### p < 0.001, ## p < 0.01, # p < 0.05, vs. control group, ∗∗∗ p < 0.001, ∗∗ p < 0.01, ∗ p < 0.05, vs. ox-LDL group.

Article Snippet: Ginsenoside Re, (Cat. B21055 , Shanghai Yuanye Biotechnology Co., Ltd, purity≥ 98 %); Simvastatin tablets (Cat. 20210925, Shandong Xinqi Pharmaceutical Co., Ltd); PFKFB3 antibody (Cat. D7H4Q, Cell Signaling Technology, Inc.); HIF-1α Rabbit pAb (Cat. A11945, ABclonal Biotechnology Co., Ltd); HK2 polyclonal antibody (Cat. 22029-1-AP, Wuhan Sanying Biotechnology Co., Ltd); vascular endothelial cadherin (VE-cadherin) antibody (Cat. 2158S, Cell Signaling Technology, Inc.); VEGFA Rabbit mAb (Cat. ab214424, abcam); VEGFR2 Rabbit mAb (Cat. 2479, Cell Signaling Technology, Inc.).

Techniques: Migration, CCK-8 Assay, Wound Healing Assay, Western Blot, Control

Journal: Journal of Ginseng Research

Article Title: Ginsenoside Re regulates PFKFB3-mediated glycolysis to inhibit endothelial cell migration to ameliorate atherosclerosis

doi: 10.1016/j.jgr.2025.11.012

Figure Lengend Snippet: Re inhibits endothelial cell migration via the PFKFB3-HIF-1α/VEGFA-VEGFR2 signaling pathways. (A) Western blot assay and quantitative analysis of PFKFB3 in HUVECs, n = 3. (B) Cell viability of PFKFB3 overexpressing ECs measured by CCK-8 assay, n = 6. (C) Western blot assay and quantitative analysis of PFKFB3 in HUVECs, n = 3. (D–E) Representative images of HUVEC induced by ox-LDL for 0 h and 12 h in wound healing experiments (bar = 50 μm) and quantification of EC migration, n = 3. (F, G) Western blot assay and quantitative data of VE-cadherin, HIF-1α, VEGFA, and VEGFR2 in HUVECs, n = 3. ### p < 0.001, ## p < 0.01, # p < 0.05, vs. control group; ∗∗∗ p < 0.001, ∗∗ p < 0.01, ∗ p < 0.05, vs. ox-LDL group.

Article Snippet: Ginsenoside Re, (Cat. B21055 , Shanghai Yuanye Biotechnology Co., Ltd, purity≥ 98 %); Simvastatin tablets (Cat. 20210925, Shandong Xinqi Pharmaceutical Co., Ltd); PFKFB3 antibody (Cat. D7H4Q, Cell Signaling Technology, Inc.); HIF-1α Rabbit pAb (Cat. A11945, ABclonal Biotechnology Co., Ltd); HK2 polyclonal antibody (Cat. 22029-1-AP, Wuhan Sanying Biotechnology Co., Ltd); vascular endothelial cadherin (VE-cadherin) antibody (Cat. 2158S, Cell Signaling Technology, Inc.); VEGFA Rabbit mAb (Cat. ab214424, abcam); VEGFR2 Rabbit mAb (Cat. 2479, Cell Signaling Technology, Inc.).

Techniques: Migration, Protein-Protein interactions, Western Blot, CCK-8 Assay, Control

Journal: bioRxiv

Article Title: Kindlin-2-Moesin interaction orchestrates sprouting angiogenesis via modulating endothelial membrane mechanics and VEGF signaling

doi: 10.64898/2026.02.24.707842

Figure Lengend Snippet: (A) HUVECs were infected with Ad-Moesin WT or Ad-Moesin T558D for 48 hrs, and total RNA was harvested for RNA-seq. The differentially expressed genes were analyzed with Gene Ontology Enrichment Analysis. The top nine gene sets are shown. Data were analyzed from GSE302986. (B) HUVECs transfected with siCtrl or si KINDLIN-2 were serum-starved (2% FBS) for 8 hrs, then stimulated with 2 μg/mL Alexa 594-labeled VEGF (red) for 30 mins. VEGF accumulation was visualized by confocal microscopy. Actin, phalloidin (green); nuclei, DAPI (blue). (C) Quantification of Alexa 594-labeled VEGF spots normalized to cellular area (n= 4 independent experiments). (D) HUVECs infected with Ad-Moesin WT or Ad-Moesin T558D were starved and stimulated with Alexa 594-labeled VEGF as in (B). VEGF accumulation was imaged by confocal microscopy. (E) Quantification of Alexa 594-labeled VEGF spots normalized to cellular area (n=3 independent experiments). (F) HUVECs transfected with siCtrl or si KINDLIN-2 were serum-starved for 8 hrs, stimulated with 50 ng/ml VEGF for 30 mins, fixed, and stained for VEGFR2 (green). Representative images show intracellular VEGFR2 vesicles. (G) Quantification of VEGFR2 vesicle number per cell from (F) (n=3 independent experiments). (H) HUVECs infected with Ad-Moesin WT or Ad-Moesin T558D were treated as in (F), and stained for VEGFR2. (I) Quantification of VEGFR2 vesicle number per cell from (H) (n= 3 independent experiments). (J) Schematic of the cell surface biotinylation assay. HUVECs were starved for 8 hrs, labeled with EZ-Link Sulfo-NHS-SS-Biotin (0.25 mg/mL) at 4 °C for 1 hr, then stimulated with VEGF (50 ng/mL) for 30 mins. After surface biotin was stripped with GSH elution buffer, total proteins were extracted. Biotinylated internalized VEGFR2 was pulled down using streptavidin magnetic beads and analyzed by western blotting. (K) Cell surface biotinylation assay for VEGFR2 internalization in siCtrl- and si KINDLIN-2 –transfected HUVECs. Input lysates show VEGFR2, Kindlin-2, and GAPDH. “Surf” represents surface VEGFR2 prior to VEGF stimulation and biotin stripping. (L) Quantification of internalized VEGFR2 normalized to surface VEGFR2 levels (n=4 independent experiments). (M) Schematic of in vivo Alexa 594-labeled VEGF uptake assay in Kindlin-2 WT and Kindlin-2 iΔEC(Pdgfb) pups. Tamoxifen was administered from P1 to P3; Alexa 594-labeled VEGF was injected intravitreally at P6 and analyzed after 30 mins. (N) Representative confocal images of the retinal angiogenic front showing uptake of Alexa 594-labeled VEGF by ECs from Kindlin-2 WT and Kindlin-2 iΔEC(Pdgfb) pups. (O) Quantification of internalized Alexa 594-VEGF at the angiogenic front (n = 4/4 pups). (P) Western blot of HUVECs transfected with siCtrl or si KINDLIN-2 , starved for 8 hrs, then stimulated with 50 ng/mL VEGF. (Q) Quantification of p-ERK levels at 5 mins after VEGF stimulation from (P) (n = 4 independent experiments). (R) Western blot of HUVECs infected with Ad-Vector, Ad-Moesin WT , or Ad-Moesin T558D , treated as in (P). (S) Quantification of p-ERK levels at 5 mins after VEGF stimulation from (R) (n = 3 independent experiments). Data are presented as mean ± SEM. * P < 0.05; ** P < 0.01; *** P < 0.001 by two-tailed Student’s t-test or one-way ANOVA followed by Tukey’s multiple comparisons test. Scale bars: 5 μm in (B), (D), (F), (H), and (N).

Article Snippet: Samples were then blocked in PBS containing 2% BSA and 0.1% Triton X-100 for 1 hr at room temperature, followed by staining with primary antibody VEGFR2 (Cell Signaling Technology, #2479S, 1:200) and the appropriate secondary antibody.

Techniques: Infection, RNA Sequencing, Transfection, Labeling, Confocal Microscopy, Staining, Cell Surface Biotinylation Assay, Magnetic Beads, Western Blot, Stripping Membranes, In Vivo, Injection, Plasmid Preparation, Two Tailed Test

Journal: bioRxiv

Article Title: The LIF-LIFR Axis Promotes Liver Regeneration via Modulation of Angiogenesis and HGF Release from LSECs

doi: 10.64898/2026.02.24.707802

Figure Lengend Snippet: (A) Volcano plot of differentially secreted proteins from primary LSECs treated with 40 ng/mL LIF versus control (n = 4). (B&C) qPCR of lifr (B) and hgf (C) levels in LSECs isolated from lifr f/f mice with indicated treatments. (D) HGF concentrations in conditioned medium from LSECs treated with hLIF at the indicated doses. (E) HGF concentrations in conditioned medium from LSECs treated with 40 ng/mL hLIF in the presence of the STAT3 inhibitor C188-9 at the indicated doses. (F) HGF concentrations in conditioned medium from LSECs isolated from lifr f/f or lifrΔEC mice treated with hLIF, hCT-1 or hOSM. (G) Western blot analysis of hepatocytes co-cultured with LSECs with or without 40 ng/mL LIF. Quantification is shown in Figure S4C. (H) Model depicting LIF action on LSECs to stimulate angiogenesis and HGF release, thereby supporting hepatocyte proliferation via paracrine regulation.

Article Snippet: The following primary antibodies were used in this study : LIFR (Proteintech, 22779-1-AP, 1:1,000 dilution), Tubulin (Proteintech, 66031-1-Ig, 1:2,000 dilution), Actin (Proteintech, 66009-1-Ig, 1:10,000 dilution), PCNA (Abcam, AB220208, 1:2,000 dilution), p-Y1175-VEGFR2 (Cell Signaling Technology, 3770S, 1:2000 dilution), Total-VEGFR2 (Cell Signaling Technology, 2479S, 1:2000 dilution), p-T705-STAT3 (Cell Signaling Technology, 9145S, 1:2000 dilution), Total-STAT3 (Cell Signaling Technology, 30835S, 1:2,000 dilution), p-T202/Y204-ERK (Cell Signaling Technology, 4370, 1:2,000 dilution) and Total-ERK (Cell Signaling Technology, 4695, 1:2,000 dilution), p-S473-AKT (Cell Signaling Technology, 4060, 1:2,000 dilution), Total-AKT (Cell Signaling Technology, 2920, 1:2000 dilution)

Techniques: Control, Isolation, Western Blot, Cell Culture