primary human retinal endothelial cells hrecs  (Innoprot Inc)

Journal: Frontiers in Immunology

Article Title: SARS−CoV−2 spike S1-mediated HIF−2α activation in retinal endothelial cells suggests a mechanism contributing to post−COVID endothelial dysfunction

doi: 10.3389/fimmu.2026.1770758

Figure Lengend Snippet: S1 induces VEGF mRNA expression in HRECs but not immune activation markers. HRECs were mock-treated (control, n=4), stimulated with S1 (100 ng/mL, n=4), or LPS (100 ng/mL, n=4) for 4 h. mRNA expression of (A) VEGF , (B) IL-6 , (C) TNF , (D) IL-8 , (E) MCP-1 , (F) CXCL1 , (G) ICAM-1 , and (H) CXCL10 was quantified by RT-qPCR and normalized to GAPDH . Each dot represents one independent experiment. A p-value of <0.05 was considered statistically significant. P-values were determined by one-way ANOVA followed by Tukey’s post hoc test (A, B, D, F-H) or Kruskal– Wallis test followed by Dunn’s post hoc test (C, E) .

Article Snippet: Primary human retinal endothelial cells (HRECs) were purchased from Innoprot and cultured in endothelial basal medium supplemented with 5% fetal bovine serum, 1% endothelial cell growth supplement, and 1% penicillin/streptomycin solution (all from Innoprot).

Techniques: Expressing, Activation Assay, Control, Quantitative RT-PCR

Journal: Frontiers in Immunology

Article Title: SARS−CoV−2 spike S1-mediated HIF−2α activation in retinal endothelial cells suggests a mechanism contributing to post−COVID endothelial dysfunction

doi: 10.3389/fimmu.2026.1770758

Figure Lengend Snippet: S1 induces high HIF-1/2α nuclear translocation and VEGFR2 upregulation in HRECs. HRECs were mock-treated (control), stimulated with S1 (100 ng/mL), or treated with CoCl 2 (100 µM) for immunofluorescence analysis. (A) HIF-1α nuclear translocation after 8 h. (B) PDK-1, BNIP-3, and GLUT-1 expression after 24 h. (C) HIF-2α nuclear translocation after 24 h and 72 h. (D) VEGFR2 expression after 24 h or 72 h. All the primary antibodies were labelled with FITC (green), and the nuclei were counterstained with DAPI (blue). Images (left) were acquired at 20× magnification, and scale bars represent 20 µm (A, D) or 100 µm (B, C) . Graphs (right) illustrate the percentage of nuclear translocation (A, C) , the corrected total cell fluorescence (B) , and the percentage of positive cells (D) . Data are represented as means ± SD. Each dot represents one independent experiment. A p-value of <0.05 was considered statistically significant. P-values were determined by one-way ANOVA followed by Tukey’s post hoc test.

Article Snippet: Primary human retinal endothelial cells (HRECs) were purchased from Innoprot and cultured in endothelial basal medium supplemented with 5% fetal bovine serum, 1% endothelial cell growth supplement, and 1% penicillin/streptomycin solution (all from Innoprot).

Techniques: Translocation Assay, Control, Immunofluorescence, Expressing, Fluorescence

Journal: Frontiers in Immunology

Article Title: SARS−CoV−2 spike S1-mediated HIF−2α activation in retinal endothelial cells suggests a mechanism contributing to post−COVID endothelial dysfunction

doi: 10.3389/fimmu.2026.1770758

Figure Lengend Snippet: S1 and plasma from PCS patients influence ROS production, impair NO availability, and disrupt barrier integrity in HRECs, effects improved by belzutifan. (A) HRECs were mock-treated (control) or stimulated with S1 (100 ng/ml) for 0–6 h, and cellular ROS levels were measured using DCFDA/H 2 DCFDA (n = 3 independent experiments). (B) HRECs were mock-treated (control) or stimulated with S1 (100 ng/ml) for 4 h, and mitochondrial ROS production was measured by flow cytometric analysis using MitoSox Red (n = 4 independent experiments). (C, D) HRECs were mock-treated (control), stimulated with S1 (100 ng/mL) or CoCl 2 (100 µM), and treated with belzutifan (50 nM) for 72 h. Immunofluorescence staining was performed for F-actin (C, red ) and VE-cadherin (D, green ) , with nuclei counterstained with DAPI (blue). Images (left) were acquired at 20× magnification, and scale bars represent 100 µm. Graphs (right) illustrate the percentage of positive cells (C) and the corrected total cell fluorescence (CTCF) (D) (n = 3 independent experiments). (E) HRECs were cultured at confluence on ECIS electrodes and then stimulated with 100 ng/mL S1 or left untreated in the presence or absence of 50 nM belzutifan for 0–72 h. The loss of barrier integrity was determined by transendothelial electrical resistance (TEER). Values were normalized to time = 0 for easier comparisons (n = 3 independent experiments). (F) HRECs were treated with 2% plasma from healthy individuals (HC, n=8) or PCS patients (n=13) for 0–6 h, and cellular ROS levels were measured using DCFDA/H 2 DCFDA. (G) Mitochondrial ROS production in HRECs exposed to 2% plasma from HC (n=8) or PCS patients (n=13) for 4 h, measured by flow cytometric analysis using MitoSox Red. (H) Total NO levels in HRECs exposed to 2% plasma from HC (n=8) or PCS patients (n=13) for 4 h and 24 h, measured using a fluorometric assay for total nitrite/nitrate levels. (I) HRECs were cultured at confluence on ECIS electrodes and exposed to 2% plasma from HC or PCS patients in the presence or absence of 50 nM belzutifan for 0–48 h. The loss of barrier integrity was determined by transendothelial electrical resistance (TEER). Values were normalized to time = 0 for easier comparisons. Data are represented as means ± SD. Each dot represents one independent experiment for S1 studies or one individual donor for plasma studies. A p-value of <0.05 was considered statistically significant. P-values were determined by two-way ANOVA followed by Tukey’s post hoc test (A, E, F, I) , Mann–Whitney U test (B) , one-way ANOVA followed by Tukey’s post hoc test (C, D) , Student’s t-test (G) , and Kruskal–Wallis test followed by Dunn’s post hoc test (H) . .

Article Snippet: Primary human retinal endothelial cells (HRECs) were purchased from Innoprot and cultured in endothelial basal medium supplemented with 5% fetal bovine serum, 1% endothelial cell growth supplement, and 1% penicillin/streptomycin solution (all from Innoprot).

Techniques: Clinical Proteomics, Control, Immunofluorescence, Staining, Fluorescence, Cell Culture, MANN-WHITNEY

Journal: World Journal of Diabetes

Article Title: Effect of ranibizumab on diabetic retinopathy via the vascular endothelial growth factor/STAT3/glial fibrillary acidic protein pathway

doi: 10.4239/wjd.v16.i5.99473

Figure Lengend Snippet: Effect of ranibizumab on the viability of adult retinal pigment epithelial 19 cells and human retinal microvascular endothelial cells. Cell viability was assessed using the Cell Counting Kit-8 assay ( n = 3, independent experiments). A: Effect of ranibizumab (0 mg/mL, 0.0625 mg/mL, 0.125 mg/mL, or 025 mg/mL) treatment on adult retinal pigment epithelial 19 (ARPE-19) cell viability; B: Effect of ranibizumab (0 mg/mL, 0.0625 mg/mL, 0.125 mg/mL, or 025 mg/mL) treatment on human retinal microvascular endothelial cell (HRMEC) viability. All results are expressed as the mean ± SD. d P < 0.0001. 1 P vs 24 hour group. 2 P vs 48 hour group. NC: Untreated group.

Article Snippet: Human retinal microvascular endothelial cells (HRMECs) and primary human retinal endothelial cells were purchased from ScienCell (Carlsbad, CA, United States) and cultured in a low-glucose (5.5 mmol/L) primary endothelial cell culture medium (ScienCell, Wuhan, Hubei Province, China) in a humidified atmosphere containing 50 mL/L carbon dioxide at 37 °C.

Techniques: Cell Counting

Journal: World Journal of Diabetes

Article Title: Effect of ranibizumab on diabetic retinopathy via the vascular endothelial growth factor/STAT3/glial fibrillary acidic protein pathway

doi: 10.4239/wjd.v16.i5.99473

Figure Lengend Snippet: mRNA expression of vascular endothelial growth factor, interleukin 6, cluster of differentiation 18, intercellular adhesion molecule, tumor necrosis factor alpha, and signal transducer and activator of transcription 3 in adult retinal pigment epithelial 19 cells and human retinal microvascular endothelial cells. Untreated (NC) (5.5 mmol/L glucose), NC + ranibizumab (5.5 mmol/L glucose + 0.125 mg/mL ranibizumab), high glucose (Hg) (25 mmol/L glucose), and Hg + ranibizumab (25 mmol/L glucose + 0.125 mg/mL ranibizumab). A: Ratios of the mRNA expression of vascular endothelial growth factor (VEGF), interleukin 6 (IL-6), cluster of 18 differentiation (CD18), intercellular adhesion molecule (ICAM), tumor necrosis factor alpha (TNF-α), and signal transducer and activator of transcription 3 (STAT3) in different adult retinal pigment epithelial 19 (ARPE-19) groups; B: Ratios of mRNA expression of VEGF, IL-6, CD18, ICAM, TNF-α, and STAT3 in different human retinal microvascular endothelial cell (HRMEC) groups. All results are expressed as the mean ± SD. a P < 0.05. b P < 0.01. 1 P vs NC group. 2 P vs Hg group.

Article Snippet: Human retinal microvascular endothelial cells (HRMECs) and primary human retinal endothelial cells were purchased from ScienCell (Carlsbad, CA, United States) and cultured in a low-glucose (5.5 mmol/L) primary endothelial cell culture medium (ScienCell, Wuhan, Hubei Province, China) in a humidified atmosphere containing 50 mL/L carbon dioxide at 37 °C.

Techniques: Expressing

Journal: World Journal of Diabetes

Article Title: Effect of ranibizumab on diabetic retinopathy via the vascular endothelial growth factor/STAT3/glial fibrillary acidic protein pathway

doi: 10.4239/wjd.v16.i5.99473

Figure Lengend Snippet: Expression of glial fibrillary acidic protein, signal transducer and activator of transcription 3 (STAT3), and phosphorylated STAT3 proteins in adult retinal pigment epithelial 19 cells and human retinal microvascular endothelial cells. Untreated (NC) (5.5 mmol/L glucose), NC + ranibizumab (5.5 mmol/L glucose + 0.125 mg/mL ranibizumab), high glucose (Hg) (25 mmol/L glucose), and Hg + ranibizumab (25 mmol/L glucose + 0.125 mg/mL ranibizumab). A: Expression of glial fibrillary acidic protein (GFAP), signal transducer and activator of transcription 3 (STAT3), and phosphorylated STAT3 (pSTAT3) proteins in adult retinal pigment epithelial 19 (ARPE-19) cells were detected by Western blot analysis; B: Expression of GFAP, STAT3, and pSTAT3 proteins in human retinal microvascular endothelial cells (HRMECs) were detected by Western blot analysis; C: Ratios of GFAP, STAT3, and pSTAT3 protein expression levels in different groups of ARPE-19 cells; D: Ratios of GFAP, STAT3, and pSTAT3 protein expression levels in different groups of HRMECs. All results are expressed as the mean ± SD. a P < 0.05. b P < 0.01. 1 P vs NC group. 2 P vs Hg group.

Article Snippet: Human retinal microvascular endothelial cells (HRMECs) and primary human retinal endothelial cells were purchased from ScienCell (Carlsbad, CA, United States) and cultured in a low-glucose (5.5 mmol/L) primary endothelial cell culture medium (ScienCell, Wuhan, Hubei Province, China) in a humidified atmosphere containing 50 mL/L carbon dioxide at 37 °C.

Techniques: Expressing, Western Blot

Journal: World Journal of Diabetes

Article Title: Effect of ranibizumab on diabetic retinopathy via the vascular endothelial growth factor/STAT3/glial fibrillary acidic protein pathway

doi: 10.4239/wjd.v16.i5.99473

Figure Lengend Snippet: Ratio between the endothelial cell to pericyte ratio and the number of acellular strands. A: Ratio between the endothelial cell to pericyte (E/P) ratio and the number of acellular strands in each group revealed by retinal periodic acid-Schiff staining (magnification: 400 ×; scale bar: 2.5 μm); B and C: Ratio between E/P and the number of acellular strands during the sixth, eighth, and tenth weeks. The yellow arrows indicate the acellular strands, while the orange arrows indicate neovascularization bud. All results are expressed as the mean ± SD. a P < 0.05. b P < 0.01. 1 P vs F group. 2 P vs B group. 3 P vs D group.

Article Snippet: Human retinal microvascular endothelial cells (HRMECs) and primary human retinal endothelial cells were purchased from ScienCell (Carlsbad, CA, United States) and cultured in a low-glucose (5.5 mmol/L) primary endothelial cell culture medium (ScienCell, Wuhan, Hubei Province, China) in a humidified atmosphere containing 50 mL/L carbon dioxide at 37 °C.

Techniques: Staining

Journal: World Journal of Diabetes

Article Title: Effect of ranibizumab on diabetic retinopathy via the vascular endothelial growth factor/STAT3/glial fibrillary acidic protein pathway

doi: 10.4239/wjd.v16.i5.99473

Figure Lengend Snippet: mRNA expression of vascular endothelial growth factor, interleukin 6, cluster of differentiation 18, intercellular adhesion molecule, tumor necrosis factor-alpha, and signal transducer and activator of transcription 3 in the retinal tissues of Sprague-Dawley rats. A: Ratio of vascular endothelial growth factor (VEGF) mRNA expression in the retinal tissue of each group of Sprague-Dawley (SD) rats; B: Ratio of interleukin 6 (IL-6) mRNA expression in the retinal tissue of each group of SD rats; C: Ratio of cluster of differentiation 18 (CD18) mRNA expression in the retinal tissue of each group of SD rats; D: Ratio of intercellular adhesion molecule (ICAM) mRNA expression in the retinal tissue of each group of rats; E: Ratio of tumor necrosis factor alpha (TNF-a) mRNA expression in the retinal tissue of each group of rats; F: Ratio of signal transducer and activator of transcription 3 (STAT3) mRNA expression in the retinal tissue of each group of rats. All results are expressed as the mean ± SD. a P < 0.05. b P < 0.01. 1 P vs A group. 2 P vs B group. 3 P vs C group. 4 P vs D group. 5 P vs F group.

Article Snippet: Human retinal microvascular endothelial cells (HRMECs) and primary human retinal endothelial cells were purchased from ScienCell (Carlsbad, CA, United States) and cultured in a low-glucose (5.5 mmol/L) primary endothelial cell culture medium (ScienCell, Wuhan, Hubei Province, China) in a humidified atmosphere containing 50 mL/L carbon dioxide at 37 °C.

Techniques: Expressing