Journal: Frontiers in Toxicology

Article Title: Nicotine-free electronic vape fluid stimulates angiogenic processes in vitro through ARF6-mediated oxidative stress

doi: 10.3389/ftox.2025.1699112

Figure Lengend Snippet: Activation of ARF6 by its guanine nucleotide exchange factor (ARNO) regulates angiogenic processes in endothelial cells induced by nicotine-free eVape. (a, b) HUVECs were exposed to 2% nicotine-free eVape fluid for 24 h, and the cell lysates were analysed by Western blotting for (a) ARF6 or (b) ARNO expression. The Western blotting membranes (representative blots are shown in (i) ) were quantified and normalised to the actin expression shown in (ii) . (c–f) HUVECs were treated with ARF6 inhibitor (NAV2729; 5 µM), ARNO inhibitor (Secin H3; 10 µM), or dimethyl sulfoxide (DMSO) as vehicle control at the same time as eVape exposure. (c) ROS accumulation was assessed by DCFDA incorporation (fluorescence at 485/535 nm) following 2 h of treatment. (d) Cell adhesion, (e) migration, and (f) angiogenic potential were measured following (d, e) 6 h and (f) 24 h of treatments. (g) Protein expression of (i) angiopoietin-2, (ii) EGF, (iii) endoglin, (iv) PIGF, (v) prolactin, and (vi) VEGF were measured using specific ELISA arrays following 24 h of treatment with 2% nicotine-free eVape fluid in the presence (closed circles) and absence (open circles) of Secin H3 (10 µM). The data are presented as mean ± SEM, n = 5–6. * p < 0.05 versus vehicle for eVape (0%); # p < 0.05 versus vehicle for NAV2729 and Secin H3 (DMSO).

Article Snippet: NAV2729 and Secin H3 were purchased from Tocris Bioscience (Abingdon, UK); Proteome Profiler Human angiogenesis array and enzyme-linked immunosorbent assay (ELISA) kits for human angiopoietin-2 (DANG20), endothelial growth factor (EGF; DEG00), endoglin (DNDG00), placental growth factor (PIGF; DPG00), prolactin (DY682), and vascular endothelial growth factor (VEGF; DVE00) were all obtained from R&D Systems (Abingdon, UK).

Techniques: Activation Assay, Western Blot, Expressing, Control, Fluorescence, Migration, Enzyme-linked Immunosorbent Assay

Journal: Pharmacology Research & Perspectives

Article Title: Vasoactivity of Rac GTPase , Cytohesin and Kinase Inhibitors in Renal Interlobar and Coronary Arteries Reveals Shared and Distinct Patterns of Inhibitory Effects in Vascular and Prostate Smooth Muscle Contraction

doi: 10.1002/prp2.70190

Figure Lengend Snippet: Effects of SecinH3 (30 μM) on agonist‐ and EFS‐induced contractions of renal, interlobar arteries. Contractions were induced by noradrenaline (a), phenylephrine (b) or EFS (c), 30 min after the addition of solvent (DMSO) to controls or SecinH3 (30 μM). Data are obtained from n = 5 independent experiments per panel, performed with arteries from n = 5 animals, with each single experiment including a control and SecinH3 group with tissues from the same animal, resulting in paired samples. Shown are means ± SD from all experiments in concentration and frequency response curves, and all single E max , EC 50 , and EF 50 values from all experiments (calculated by curve fitting).

Article Snippet: EHT1864, NSC23766, SR7826, LIMKi3, CMPD101, FRAX486, and SecinH3 were obtained from Tocris (Bristol, UK).

Techniques: Solvent, Control, Concentration Assay

Journal: Pharmacology Research & Perspectives

Article Title: Vasoactivity of Rac GTPase , Cytohesin and Kinase Inhibitors in Renal Interlobar and Coronary Arteries Reveals Shared and Distinct Patterns of Inhibitory Effects in Vascular and Prostate Smooth Muscle Contraction

doi: 10.1002/prp2.70190

Figure Lengend Snippet: Effects of SecinH3 on cholinergic contractions of coronary arteries. Contractions were induced 30 min after addition of solvent (DMSO) to controls or 30 μM SecinH3 (a, b) or 10 μM SecinH3 (c, d) by carbachol (a, c) or methacholine (b, d). Data are obtained from n = 5 independent experiments per panel, performed with arteries from n = 5 animals, with each single experiment including a control and SecinH3 group with tissues from the same animal, resulting in paired samples. Shown are means ± SD from all experiments in concentration response curves together with p values from two‐way ANOVA for whole groups, and all single E max and EC 50 values from all experiments (calculated by curve fitting) analyzed by Student's t ‐test. p values ≥ 0.05 are not shown. E max values marked gray represent maximum tensions in concentration response curves, as curve fitting was not possible in these experiments, and EC 50 values marked in gray have been estimated from concentration response curves in these experiments.

Article Snippet: EHT1864, NSC23766, SR7826, LIMKi3, CMPD101, FRAX486, and SecinH3 were obtained from Tocris (Bristol, UK).

Techniques: Solvent, Control, Concentration Assay

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: The endocannabinoid 2-arachidonoylglycerol is released and transported on demand via extracellular microvesicles.

doi: 10.1073/pnas.2421717122

Figure Lengend Snippet: Fig. 4. 2-AG is specifically sorted into microvesicles in a DAGL- and Arf6-dependent process. (A and B) Fold-enrichment (ATP/Vehicle) of lipids in EVs (A) or cells (B) following vehicle (MQ) or ATP-treatment for 30 min at 37 °C. (C) Size distribution of EVs determined by nanoparticle tracking analysis (NTA) (n = 3 videos). (D) Representative western blot and (E) quantification of CD9 signal intensity in EVs (n = 9). (F and G) 2-AG levels in EVs (F) and cells (G) relative to vehicle-treated control (n = 10/7 DMSO/DH376). (H) Size distribution of EVs determined by nanoparticle tracking analysis (NTA) (n = 3 videos). (I) Representative western blot and (J) quantification of CD9 signal intensity in EVs (n = 7). (K and L) 2-AG levels in EVs (K) and cells (L) relative to vehicle-treated control (n = 3). Cells were treated with 1 µM DH376 (C–G) or 10 µM SecinH3 (H–L) for 20 min prior to vehicle (MQ) or ATP-treatment for 30 min at 37 °C, followed by EV isolation. Data are shown as mean ± SD. n is individual biological replicates/EV isolations. Statistical testing was performed using one-way ANOVA with Tukey’s correction for multiple comparisons. *P < 0.05, **P < 0.01, ***P < 0.001, ns not significant P > 0.05.

Article Snippet: SecinH3 (S7685), ML- 7 (S8388), GW4896 (S7609), and Sotrastaurin (S2791) were purchased from Selleck Chemicals.

Techniques: Western Blot, Control, Isolation