Journal: Angiogenesis

Article Title: Exploiting porphyrin metabolism to inhibit angiogenesis

doi: 10.1007/s10456-026-10034-y

Figure Lengend Snippet: ALA promotes porphyrin overdrive in human microvascular EC a Intracellular porphyrins levels in HMEC treated with 5 mM ALA for 4, 24, and 72 h. b – c Fluorescence imaging and relative quantification of intracellular porphyrins (magenta) in HMEC treated with 5 mM ALA and controls. Scale Bar: 50 µm. n = 12 d Intracellular heme levels in HMEC treated with 5 mM ALA for 4, 24, and 72 h. e Mitochondrial activity of ALAS in HMEC treated with 5 mM ALA for 24 h (hrs). f – h Representative Western Blot images ( f ) and their quantification g – h of ALAS1 and HO-1 protein levels in HMEC treated with 5 mM ALA for 24 and 72 h. i – j Differential expression of heme-related genes in HMEC upon 24 h of 5 mM ALA treatment. The heatmap shown in ( i ) displays differences in gene expression levels, represented as fold change (FC) respect to not-treated (NT) cells. Volcano plot shown in ( j ) represents the differences in gene expression levels versus -log10(q value). The dotted line indicates the significance threshold of FDR q < 1% (-log10(q value) = 2). The plotted results represent mean values obtained from at least 5 individual biological replicates. k – l Extracellular levels of porphyrins ( k ) and heme ( l ) in HMEC after 4, 24, and 72 h of 5 mM ALA supplementation. Data are expressed as mean ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. For statistical analyses, ordinary one-way ANOVA test with Tukey’s multiple comparisons ( a , d , g , h , k , l ) and parametric unpaired t test ( c , e ) were used. FC, fold change; NT, not treated controls; hrs, hours; ALA, 5-amminolevulinic acid

Article Snippet: Human adult dermal microvascular endothelial cells (HMEC-1, RRID:CVCL_0307) were purchased by ATCC, propagated in MCDB131 (Thermo Fisher Scientific, Waltham, MA USA, catalog n°10,372,019) supplemented with 10% heat-inactivated low-endotoxin FBS (GIBCO by Thermofisher Scientific, Waltham, MA USA, catalog n10270106), 10 mM GlutaMAXTM Supplement (Thermo Fisher Scientific, Waltham, MA USA, catalog n°35,050,061), 10 ng/mL Epidermal Growth Factor (Thermo Fisher Scientific, Waltham, MA USA, catalog n° PHG0315), 1 μg/mL Hydrocortisone-Water Soluble (Sigma Aldrich, St. Louis, MO USA, catalog n° H0396) and used up to passage 12.

Techniques: Fluorescence, Imaging, Quantitative Proteomics, Activity Assay, Western Blot, Gene Expression

Journal: Angiogenesis

Article Title: Exploiting porphyrin metabolism to inhibit angiogenesis

doi: 10.1007/s10456-026-10034-y

Figure Lengend Snippet: ALA inhibits in vitro angiogenesis on human microvascular EC ( a and b ) Proliferation of HMEC treated with increasing concentrations of ALA (500 nM, 0.1 mM, 5 mM) at various time points. Scale bar: 500 µm. n = 6 (c and d) Wound healing experiment of HMEC treated with increasing concentrations of ALA (500 nM, 0.1 mM, 5 mM) at various time points. Scale bar: 500 µm. n > 8 (e – h) in vitro tubulogenesis assay performed on 5 mM ALA-treated and control HMEC. Quantification of the total length ( f ) of the networks, number of nodes ( g ) and number of branches ( h ) are shown. Scale bar: 500 µm. Data are expressed as mean ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. For statistical analyses ordinary two-way ANOVA test with Tukey’s multiple comparisons ( b , d ) and parametric unpaired t-test ( f – h ) were used. FC, fold change; NT, not treated controls; hrs, hours; ALA, 5-amminolevulinic acid

Article Snippet: Human adult dermal microvascular endothelial cells (HMEC-1, RRID:CVCL_0307) were purchased by ATCC, propagated in MCDB131 (Thermo Fisher Scientific, Waltham, MA USA, catalog n°10,372,019) supplemented with 10% heat-inactivated low-endotoxin FBS (GIBCO by Thermofisher Scientific, Waltham, MA USA, catalog n10270106), 10 mM GlutaMAXTM Supplement (Thermo Fisher Scientific, Waltham, MA USA, catalog n°35,050,061), 10 ng/mL Epidermal Growth Factor (Thermo Fisher Scientific, Waltham, MA USA, catalog n° PHG0315), 1 μg/mL Hydrocortisone-Water Soluble (Sigma Aldrich, St. Louis, MO USA, catalog n° H0396) and used up to passage 12.

Techniques: In Vitro, Control

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