Journal: Redox Biology

Article Title: Human lung microvascular endothelial cell protein modification by 2-chlorohexadecanoic acid: RhoA mediates 2-chlorohexadecanoic acid-elicited endothelial activation

doi: 10.1016/j.redox.2025.103596

Figure Lengend Snippet: 2-ClHyA modified HLMVEC proteins . (A) Venn diagram of the 2-ClHyA- and HyA-modified proteins in HLMVECs. (B) Protein class distributions of the 11 proteins identified exclusively in the 2-ClHyA sample group, analyzed using PANTHER classification system. (C) Protein class distributions of the 194 proteins identified in both the 2-ClHyA and HyA sample groups, analyzed using PANTHER.

Article Snippet: Human lung microvascular endothelial cells (HLMVEC) (PromoCell, C‐12281, Heidelberg, Germany) were cultured at 37 °C in EGM‐2MV medium (Sigma, cat. CC‐3202), in a humidified atmosphere with 95 % air/5 % CO 2 (v/v).

Techniques: Modification

Journal: Redox Biology

Article Title: Human lung microvascular endothelial cell protein modification by 2-chlorohexadecanoic acid: RhoA mediates 2-chlorohexadecanoic acid-elicited endothelial activation

doi: 10.1016/j.redox.2025.103596

Figure Lengend Snippet: Bioinformatic characterization of HLMVEC proteins modified by 2-ClHyA and HyA: Enriched Gene Ontology (GO) cellular components are shown, and were the only GO categories enriched in the 2-ClHyA modified proteome. Over-representation analysis was performed with the A.GO. TOOL using Bonferroni p < 0.05 for computing FDR with a p value cutoff of 0.01. Over-representation was compared to the HLMVEC proteome as a background. S value is a combination of -log p value and effect size. Effect size represents the difference in proportions of the foreground and background proteomes.

Article Snippet: Human lung microvascular endothelial cells (HLMVEC) (PromoCell, C‐12281, Heidelberg, Germany) were cultured at 37 °C in EGM‐2MV medium (Sigma, cat. CC‐3202), in a humidified atmosphere with 95 % air/5 % CO 2 (v/v).

Techniques: Modification

Journal: Redox Biology

Article Title: Human lung microvascular endothelial cell protein modification by 2-chlorohexadecanoic acid: RhoA mediates 2-chlorohexadecanoic acid-elicited endothelial activation

doi: 10.1016/j.redox.2025.103596

Figure Lengend Snippet: Protein-protein association network of the proteins modified exclusively by 2-ClHyA in HLMVEC. A) Association network of all proteins modified by 2-ClHyA (205 proteins). Red nodes indicate proteins identified in the cell-cell junction family of the Subcellular location group. Blue nodes indicate proteins identified in the actin filament-based process family of the Biological Process (GO) group. Green nodes indicate proteins identified in the proteasomal protein catabolic process family of the Biological Process (GO) group. Yellow nodes indicate proteins identified in the generation of precursor metabolites and energy family of the Biological Process (GO) group. Network generated using STRING, with confidence set to medium. B) Association network of proteins exclusively modified by 2-ClHyA (11 proteins). Blue nodes indicate proteins identified in the tight junction family of the KEGG Pathways. Network generated using STRING, with confidence set to medium. RhoA is highlighted in a red rectangle in STRING networks in both A and B. C) List of the 11 proteins exclusively modified by 2-ClHyA. PSM denotes peptide spectrum map.

Article Snippet: Human lung microvascular endothelial cells (HLMVEC) (PromoCell, C‐12281, Heidelberg, Germany) were cultured at 37 °C in EGM‐2MV medium (Sigma, cat. CC‐3202), in a humidified atmosphere with 95 % air/5 % CO 2 (v/v).

Techniques: Modification, Generated

Journal: Redox Biology

Article Title: Human lung microvascular endothelial cell protein modification by 2-chlorohexadecanoic acid: RhoA mediates 2-chlorohexadecanoic acid-elicited endothelial activation

doi: 10.1016/j.redox.2025.103596

Figure Lengend Snippet: RhoA mediated 2-ClHA-elicited HLMVEC barrier leak. A) HLMVECs were grown on ECIS arrays to confluency and then were either pre-treated with C3 or vehicle (arrow 1) 4h prior to lipid addition, Rhosin or vehicle (arrow 2) 2h prior to lipid addition. Treatment with 2-ClHA (10 μM) or vehicle is indicated by arrow 3. Resistance is normalized to levels measured 30 min prior to starting pretreatment. Data are mean±S.E.M.; n = 20. B) Individual data points following 6h of treatment conditions in panel A are shown as well as other conditions including Rhosin and C3 treatment without lipid treatment as well as indicated concentrations of either 2-ClHA or HA. Average change in normalized resistance at 6h following lipid treatment, data are mean±S.E.M.; n = 20 independent experiments. C) RhoA activity in HLMVEC treated with either BSA (vehicle), 2-ClHA, or HA (10 μM) was measured as described in “Materials and Methods”. HLMVECs were pretreated with C3 (1 μg/ml) or Rhosin (30 μM) 4h or 2h, respectively prior to lipid treatments. Data are mean±S.E.M.; n = 8–9 independent experiments. D) Ang-2 level in HLMVEC cell culture media was determined in cells treated with either BSA (vehicle), 2-ClHA, or HA (10 μM) was measured as described in “Materials and Methods”. HLMVECs were pretreated with C3 (1 μg/ml) or Rhosin (30 μM) 4h or 2h, respectively prior to lipid treatments. Data are mean±S.E.M.; n = 5 independent experiments. ∗, ∗∗∗, and ∗∗∗∗ indicate p < 0.05, p < 0.001, and p < 0.0001 for indicated comparisons.

Article Snippet: Human lung microvascular endothelial cells (HLMVEC) (PromoCell, C‐12281, Heidelberg, Germany) were cultured at 37 °C in EGM‐2MV medium (Sigma, cat. CC‐3202), in a humidified atmosphere with 95 % air/5 % CO 2 (v/v).

Techniques: Activity Assay, Cell Culture

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: Atrial natriuretic peptide attenuates LPS-induced lung vascular leak: role of PAK1

doi: 10.1152/ajplung.00202.2009

Figure Lengend Snippet: Effects of atrial natriuretic peptide (ANP) on endothelial cell (EC) barrier dysfunction induced by inflammatory agonists. EC were plated on gold microelectrodes and grown to confluence. At the time point indicated by the 1st arrow, cells were pretreated with ANP (100 nM, 20 min). At the time point indicated by the 2nd arrow, cells were stimulated with LPS (200 ng/ml; A and B), TNF-α (20 ng/ml; C), or a combination of IL-6 (25 ng/ml) and its soluble coreceptor, IL-6+SR (100 ng/ml; D), and transendothelial electrical resistance was monitored over time. Data are expressed as means ± SD of 3–10 independent experiments; n = 3 per condition for each experiment. HPAEC, human pulmonary macrovascular EC; HLMVEC, human lung microvascular EC.

Article Snippet: Human pulmonary macrovascular (HPAEC) and microvascular (HLMVEC) EC were obtained from Lonza (Allendale, NJ).

Techniques: