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: 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