Journal: The Journal of Biological Chemistry

Article Title: Small molecule intervention of actin-binding protein profilin1 reduces tumor angiogenesis in renal cell carcinoma

doi: 10.1016/j.jbc.2025.111033

Figure Lengend Snippet: Discovery of a novel analog of C74 with superior antiangiogenic activity in vitro and in vivo . A and B , docking models of C74 and UP-6 interaction with Pfn1—the binding modes of C74 ( panel A ) and UP-6 ( panel B ) as predicted by GNINA. Blue arrow indicates the altered structure on each compound. The unmodified hydroxypyrazoles are predicted to make an identical network of hydrogen bonds in both compounds while the modified aryl ring can potentially interact with R74 and H119. C and D , representative images ( panel C ; 4x magnification) and quantification ( panel D ; based on 4x field images) of cord formation by HmVECs on Matrigel treated with either 20 μM compound UP6 or equivalent DMSO control. Cord length is determined by the distance between each cord junction, i.e. cord intersections. Each data point represents quantification of cord formation in a single field of observation (two-tailed Student’s t test; ∗∗∗∗, p < 0.0001; n = 3 experiments; the scale bar represents 200 μm). E - F , representative fluorescence images ( panel E ) and quantification ( panel F ) of cord formation by HmVECs (labeled with cell tracker dye) subjected to 10 μM of either C74 or UP6 versus DMSO (control) (One-way ANOVA with Tukey multiple comparison post hoc test, ∗ - p < 0.05; ns – not significant; the scale bar represents 200 μm). G - H , representative images of CD31 immunohistochemistry with hematoxylin counterstaining ( panel G ; 4 × magnification) and quantification ( panel H ; based on 20X field images) of CD31+ cell infiltration in the subcutaneously implanted Matrigel plugs in Balb/C mice coinjected with a single 200 μM dosage of either C74 or UP-6 or DMSO control. Data are summarized from quantification of 10 bilaterally injected plugs (n = 5 mice) per treatment group (One-way ANOVA with Tukey post hoc test, ∗∗, p < 0.01; ∗∗∗∗, p < 0.0001; the scale bar represents 200 μm). Pfn1, profilin1; HmVEC, human microvascular EC; DMSO, dimethyl sulfoxide.

Article Snippet: Human microvascular ECs (HmVECs, ATCC, CRL-3243) were cultured in MCDB-131 media with growth supplements as described previously ( ).

Techniques: Activity Assay, In Vitro, In Vivo, Binding Assay, Modification, Control, Two Tailed Test, Fluorescence, Labeling, Comparison, Immunohistochemistry, Injection

Journal: The Journal of Biological Chemistry

Article Title: Small molecule intervention of actin-binding protein profilin1 reduces tumor angiogenesis in renal cell carcinoma

doi: 10.1016/j.jbc.2025.111033

Figure Lengend Snippet: . In vitro proof-of-concept of angiogenesis inhibition by C74 upon release by UTMD of lipid microbubbles. A and B , representative cord-formation images of HmVECs for various treatment settings ( panel A ): NT: DMSO (vehicle control); C74: treatment with unencapsulated C74 (this group shows effect of C74 alone); MB US: empty MB with UTMD (this control group tests if bubble cavitation alone causes reduction in cord formation), C74 MB: C74-encapsulated MB with no UTMD (this group demonstrates that uncavitated bubbles do not cause an effect even though they are dosed with C74); C74 MB US: C74-encapsulated MB with UTMD (this is the true test group showing that when bubbles are destroyed and C74 is released, we see reduction in cord formation), and C74 US: C74-encapsulated MB with UTMD followed by immediate washout and replacement with C74-devoid culture media (this group tests whether C74 immediately enters cells post bubble destruction). The bar graph in panel B depicts the relative cord lengths between the different treatment groups—cord length is defined as the distance between the junctional i.e. cord-intersectional points. (One-way ANOVA with Tukey post hoc test, ∗∗∗∗ p < 0.0001 from n = 3 experiments). HmVEC, human microvascular EC; UTMD, ultrasound-targeted microbubble destruction; DMSO, dimethyl sulfoxide.

Article Snippet: Human microvascular ECs (HmVECs, ATCC, CRL-3243) were cultured in MCDB-131 media with growth supplements as described previously ( ).

Techniques: In Vitro, Inhibition, Control

Journal: PLOS Neglected Tropical Diseases

Article Title: In vitro and in vivo endothelial interactions of Leptospira species are markers of virulence

doi: 10.1371/journal.pntd.0013939

Figure Lengend Snippet: (top) HMEC-1 cells were grown to post-confluence, bacteria were added at an MOI of 20 and incubated for one hour. Non-associated bacteria were removed by washing and qPCR was performed to quantify associated bacteria. Strains associate with endothelial cells to varying extents, with 3/6 P1 + strains and 2/5 P1- species binding significantly more than Lb P. (bottom) HMEC-1 cells were grown to confluence on glass coverslips and infected at an MOI of 20 for 24 h. After washing to remove unbound bacteria, cells were fixed, stained for VE-cadherin, and mounted with Prolong Diamond Antifade Mountant with DAPI. Binary area was quantified as previously described and subtracted from uninfected controls to define “VE-cadherin disruption”. P1 + Leptospira (4/6) disrupt VE-cadherin more than other clades (3/10). Spearman correlation analysis determines cell association and VE-cadherin disruption correlate with r s = 0.644 and p = 0.0085. Strains are ordered based upon presence of virulence-associated genes in their genomes . Mean ± SEM is plotted. Each column is compared to LbP . * p < 0.05, # p < 0.01, & p < 0.001, $ p < 0.0001.

Article Snippet: Human dermal microvascular endothelial cells (HMEC-1) [ ] were obtained from ATCC (CRL-3243) and cultured as described previously [ , ] in MCDB medium at 37°C with 5% CO 2 .

Techniques: Bacteria, Incubation, Binding Assay, Infection, Staining, Disruption

Journal: Journal of Extracellular Biology

Article Title: Angiogenic Potential of Small Extracellular Vesicles Produced by Stimulated Mesenchymal Stromal Cells Under Hypoxic Conditions

doi: 10.1002/jex2.70105

Figure Lengend Snippet: sEVs characterisation. sEVs were isolated by ultracentrifugation (100,000 × g ). (A) Representative size distribution of sEVs derived from MSCs (NTA) (1:500 dilution). (B) Representative size distribution of sEVs derived from SH‐MSCs (NTA) (1:500 dilution). (C) MSC sEVs (grey box, N = 14, n = 2) and SH‐MSC sEVs (blue box, N = 14, n = 2) diameters (Mann–Whitney test; ** p < 0.01). (D) Transmission electron microscopy: MSC sEVs (left), SH‐MSC sEVs (right); White arrows indicate sEVs, scale bar: 200 nm. (E) Protein content (in µg/1 × 10 9 sEVs) in MSC sEVs (grey box, N = 12) and SH‐MSC sEVs (blue box, N = 12) (Mann–Whitney test, ns). (F) sEV markers [MACSPlex (MSC sEVs n = 3, and SH‐MSC sEVs n = 3) (Mann–Whitney test, * p < 0.05); and western blot]. (G) CD31 (PECAM1) [MACSPlex (MSC sEVs n = 3, and SH‐MSC sEVs n = 3); and western blot]. HMEC‐1 were used as an ECs‐control. (H) MSC markers, CD29 (β1‐subunit integrin), and CD49e (α5‐subunit integrin) [MACSPlex (MSC sEVs n = 3, and SH‐MSC sEVs n = 3); and western blot]; (Mann–Whitney test, * p < 0.05).

Article Snippet: HMEC‐1 (Human dermal microvascular ECs‐1; ATCC CRL‐3243) were cultured in Endothelial Cell Growth Medium (Promocell) supplemented with 10% FBS and 1% antibiotic‐antimycotic.

Techniques: Isolation, Derivative Assay, MANN-WHITNEY, Transmission Assay, Electron Microscopy, Western Blot, Control

Journal: Journal of Extracellular Biology

Article Title: Angiogenic Potential of Small Extracellular Vesicles Produced by Stimulated Mesenchymal Stromal Cells Under Hypoxic Conditions

doi: 10.1002/jex2.70105

Figure Lengend Snippet: Interaction of MSC‐ and SH‐MSC‐derived sEVs with HMEC‐1 cells. (A) Scanning electron microscopy: representative experiment showing the interaction of sEVs with HMEC‐1 (after 1‐h contact, at 37°C). White arrows indicate sEVs (scale bar: 5 and 20 µm). (B) Fluorescence microscopy: internalisation (after 6 h of incubation at 37°C) of MSC sEVs and SH‐MSC sEVs by HMEC‐1. sEVs were coloured green by CellTrace Oregon Green, green arrows indicate sEVs; cells were coloured red by CellTrace Calcein Red‐Orange; Merge: Yellow‐stained EVs have been internalised and visible on orthogonal view (z‐axis). Yellow arrows indicate sEVs uptake (scale bar: 10 µm). (C) Effect of sEVs on the proliferation of HMEC‐1 cells ( N = 7, n = 6). HMEC‐1 were cultured in medium supplemented with 5% FBS sEV‐depleted [Positive (Pos) control, red box] or with 2.5% FBS sEV‐depleted [Negative (Neg) control, green box] or with 2.5% FBS sEV‐depleted in presence of two concentrations (D1 = 0.4 ± 0.08 × 10 9 /mL; D2 = 1.7 ± 1.0 × 10 9 /mL) of MSC sEVs (D1: light grey box; D2: grey box) or SH‐MSC sEVs (D1: light blue box; D2: blue box). Cell proliferation in the positive control was significantly higher (Kruskal–Wallis test, **** p < 0.0001) than negative control or than MSC sEVs (D1 and D2) or than SH‐MSC sEVs (D1). Kruskal–Wallis test was used for multiple comparisons adjustments (** p < 0.01; *** p < 0.001; **** p < 0.0001).

Article Snippet: HMEC‐1 (Human dermal microvascular ECs‐1; ATCC CRL‐3243) were cultured in Endothelial Cell Growth Medium (Promocell) supplemented with 10% FBS and 1% antibiotic‐antimycotic.

Techniques: Derivative Assay, Electron Microscopy, Fluorescence, Microscopy, Incubation, Staining, Cell Culture, Control, Positive Control, Negative Control

Journal: Journal of Extracellular Biology

Article Title: Angiogenic Potential of Small Extracellular Vesicles Produced by Stimulated Mesenchymal Stromal Cells Under Hypoxic Conditions

doi: 10.1002/jex2.70105

Figure Lengend Snippet: Quantitative measure of sEV cell uptake. Fluorescence microscopy analysis was performed at 6 h (Figure , and ) and at 24 h (S3). (A) Quantitative measurement of HMEC‐1 uptake: sEVs ( N = 4) (grey box, n = 9 at 6 h; n = 10 at 24 h) and SH‐MSC sEVs (blue box, n = 11 at 6 h; n = 17 at 24 h) (Kruskal–Wallis test, * p < 0.05; ** p < 0.01). (B) Quantitative measurement of C2C12 uptake: sEVs ( N = 2) (grey box, n = 10 at 6 h; n = 9 at 24 h) and SH‐MSC sEVs (blue box, n = 6 at 6 h; n = 8 at 24 h) (Kruskal–Wallis test, * p < 0.05; **** p < 0.0001). (C) Quantitative measurement of macrophage uptake: sEVs ( N = 4) (grey box, n = 9 at 6 h; n = 9 at 24 h) and SH‐MSC sEVs (blue box, n = 12 at 6 h; n = 17 at 24 h) from four independent experiments. (MFI: median fluorescence intensity) (Kruskal–Wallis test, not significant).

Article Snippet: HMEC‐1 (Human dermal microvascular ECs‐1; ATCC CRL‐3243) were cultured in Endothelial Cell Growth Medium (Promocell) supplemented with 10% FBS and 1% antibiotic‐antimycotic.

Techniques: Fluorescence, Microscopy

Journal: Cells

Article Title: The Role of Aldosterone in Vascular Permeability in Diabetes

doi: 10.3390/cells15010089

Figure Lengend Snippet: Permeability of the human dermal microvascular endothelial cell (HMEC-1) monolayer under normal (NORM) or hyperglycemic (HG) conditions after MR blockade; ( a ) with 15 min of aldosterone exposure, and ( b ) with 60 min of aldosterone exposure. ALDO—aldosterone; EPL—eplerenone. Results are presented as mean ± SD; n = 6–9. Statistical relationships among groups were visualized using compact letter displays, where groups sharing the same letter are not significantly different, whereas groups without a common letter differ significantly ( p < 0.05).

Article Snippet: Immortalized human dermal microvascular endothelial cells (HMEC-1; ATCC ® CRL-3243TM; ATCC, Glasgow, UK) were maintained at 37 °C in a 5% CO 2 atmosphere in MCB131 medium (GibcoTM, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum, streptomycin–penicillin (100 U/mL; Sigma-Aldrich, USA), L-glutamine (2 mM; Sigma-Aldrich, USA), hydrocortisone (1 μg/mL; Sigma-Aldrich, USA), heparin (25 U; Sigma-Aldrich, USA), and endothelial growth factor (30 μg/mL; Sigma-Aldrich, USA).

Techniques: Permeability

Journal: Cells

Article Title: The Role of Aldosterone in Vascular Permeability in Diabetes

doi: 10.3390/cells15010089

Figure Lengend Snippet: Mineralocorticoid receptor–dependent mechanisms of diabetic skin vascular permeability and their modulation by eplerenone. ( a ) Experimental diabetes and hyperglycemia are associated with increased aldosterone (ALDO) levels and reduced expression of 11β-hydroxysteroid dehydrogenase type 2 (HSD11β2) in the skin, leading to loss of mineralocorticoid receptor (MR) selectivity and enhanced MR activation by both mineralocorticoids and glucocorticoids. MR overactivation triggers genomic and non-genomic signaling pathways that promote inflammation, oxidative stress, vascular endothelial growth factor (VEGF) up-regulation, and endothelial activation characterized by increased von Willebrand factor (vWF) exocytosis. These processes converge to destabilize endothelial junctions, resulting in endothelial barrier dysfunction and increased skin microvascular permeability, which contributes to the development of diabetic skin disorders. ( b ) Pharmacological MR blockade with eplerenone (EPL) attenuates diabetes-induced skin vascular permeability. EPL reduces MR-dependent inflammatory signaling, oxidative stress, VEGF expression, and vWF exocytosis, thereby improving endothelial barrier function and limiting vascular leakage. These protective effects occur independently of changes in blood pressure or glycemic control, highlighting the pleiotropic vasculoprotective actions of EPL in diabetic skin microvasculature.

Article Snippet: Immortalized human dermal microvascular endothelial cells (HMEC-1; ATCC ® CRL-3243TM; ATCC, Glasgow, UK) were maintained at 37 °C in a 5% CO 2 atmosphere in MCB131 medium (GibcoTM, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum, streptomycin–penicillin (100 U/mL; Sigma-Aldrich, USA), L-glutamine (2 mM; Sigma-Aldrich, USA), hydrocortisone (1 μg/mL; Sigma-Aldrich, USA), heparin (25 U; Sigma-Aldrich, USA), and endothelial growth factor (30 μg/mL; Sigma-Aldrich, USA).

Techniques: Permeability, Expressing, Activation Assay, Protein-Protein interactions, Control