Journal: Journal of Molecular and Cellular Cardiology Plus

Article Title: Semaphorin 3F is elevated in serum of heart failure patients and inhibits cardiac angiogenesis via the VEGF/Akt/eNOS pathway

doi: 10.1016/j.jmccpl.2025.100470

Figure Lengend Snippet: Serum from HF patients inhibits sprouting and branching in endothelial cells, and Sema3F levels in serum from HF patients correlate with its anti-angiogenic effect HUVECs were incubated for 16 to 18 h with 30 % serum from patients with CAD ( n = 10), HF ( n = 21) or EBM with 2 % FCS (control, n = 12) and then assayed on Matrigel® for 2.5 h. (A) Cumulative sprout length and (B) branch points of capillary-like structures were diminished in response to HF serum compared to treatment with serum from CAD subjects. Box plots extend from the 25th to the 75th percentile with the median marked. Whiskers extend from the 10th to the 90th percentile. The results were analyzed using the one-way ANOVA test followed by Bonferroni's multiple comparisons test: *p < 0.05 CAD vs. controls or CAD vs. HF (n, number of patients). (C) Serum Sema3F levels were significantly higher in patients with HF (n = 21) compared to patients with CAD (n = 10). Box plots extend from the 25th to the 75th percentile with the median marked. Whiskers extend from the 10th to the 90th percentile. Mann-Whitney test, *p < 0.05 CAD vs HF. (D) The cumulative sprouting length failed to attain statistical significance (Spearman's r = 0.4180, p = 0.0666), the branch points of the Matrigel® wells that had been treated with serum from HF patients (n = 21) exhibited an inverse correlation with serum Sema3F concentration (E, Spearman's r - 0.4543, p < 0.05).

Article Snippet: Human cardiac endothelial cells (HCECs) were purchased from PELOBIOTECH GmbH, Planegg, DE and were grown at 37 °C, 5 % CO 2 in growth factor enhanced microvascular endothelial cell medium (PELOBIOTECH GmbH, Planegg, DE) supplemented with 10 % fetal calf serum (FCS) obtained from Lonza Pharma & Biotech (Verviers, BEL).

Techniques: Incubation, Control, MANN-WHITNEY, Concentration Assay

Journal: Journal of Molecular and Cellular Cardiology Plus

Article Title: Semaphorin 3F is elevated in serum of heart failure patients and inhibits cardiac angiogenesis via the VEGF/Akt/eNOS pathway

doi: 10.1016/j.jmccpl.2025.100470

Figure Lengend Snippet: Sema3F is an inhibitor of angiogenesis in Matrigel® sprouting assays. (A-C) In cardiac endothelial cells (HCECs) Sema3F abolished the pro-angiogenic effect of VEGF. HCECs were cultured with the indicated amounts of Sema3F or VEGF alone or in combination over 16 to 18 h. HCECs were then assayed on Matrigel® for further 2.5 h. (A) Representative micrographs of stimulated HCECs are shown. (B&C) Sema3F prevented VEGF-induced cumulative sprout length (B) and branching of HCECs (C). Data from 4 independent experiments were shown (n, number of biological replicates). The results were analyzed using the one-way ANOVA and Bonferroni's multiple comparisons test: * p < 0.05. Figs. D—F show the effect of Sema3F knockdown on angiogenesis of HCECs. The siRNA-based Sema3F knockdown resulted in a potentiated pro-angiogenic effect of VEGF. HCECs transfected with Sema3F-specific siRNA (siSema3F) or control siRNA (ctrl-siRNA) were treated with 100 ng/ml VEGF or EBM with 2 % FCS for 18 h before being assayed on Matrigel®. (D) Representative micrographs are shown. Sema3F knockdown led to a significant increase in cumulative sprout length (E) and number of branch points (F). (E&F) Data from 4 independent experiments were shown (n, number of biological replicates). All box plots extend from the 25th to the 75th percentile with the median marked. Whiskers extend from the 10th to the 90th percentile. The results were analyzed using one-way ANOVA followed by Bonferroni's multiple comparison test: * p < 0.05.

Article Snippet: Human cardiac endothelial cells (HCECs) were purchased from PELOBIOTECH GmbH, Planegg, DE and were grown at 37 °C, 5 % CO 2 in growth factor enhanced microvascular endothelial cell medium (PELOBIOTECH GmbH, Planegg, DE) supplemented with 10 % fetal calf serum (FCS) obtained from Lonza Pharma & Biotech (Verviers, BEL).

Techniques: Cell Culture, Knockdown, Transfection, Control, Comparison