Journal: bioRxiv

Article Title: A Biphasic Effect of Alcohol on Endothelial Plasticity Through Regulation of Endothelial-to-Mesenchymal Transition

doi: 10.64898/2026.04.14.718463

Figure Lengend Snippet: (a-c) SM22α, eNOS, αSMA, and Fibronectin (FN) mRNA levels determined by qRT-PCR in human coronary artery endothelial cells (HCAEC) following treatment with TGFβ1 (10 ng/ml, 7d), hypoxia (5% O 2 , 4d) or TGFβ1 plus hypoxia (4d), in the absence or presence of ethanol (EtOH) at concentrations indicated (0-100 mM range). (d) SM22α, eNOS, and Cdh5 mRNA levels in HCAEC treated with TGFβ1 (10 mg/ml, 2d) +/- EtOH 25 mM or 100 mM. (e) Representative western blots showing CD31, Cdh5, SM22α, and SNAIL protein expression in HCAEC treated with TGFβ1, IL-1β, or Hypoxia +/- EtOH (25 mM or 100 mM). β-actin or GAPDH were used as loading controls. Data are mean±SEM, n=3. *p<0.05 vs control (no treatment), #p<0.05 vs TGFβ1 or hypoxia.

Article Snippet: Primary antibodies used include human CD31 (Cat. # AF806, R&D Systems), αSMA (MAB1420, R&D systems), SM22α (Cat. # 36090 CST), and CDH5 (Cat. # 2500, CST).

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Control

Journal: bioRxiv

Article Title: A Biphasic Effect of Alcohol on Endothelial Plasticity Through Regulation of Endothelial-to-Mesenchymal Transition

doi: 10.64898/2026.04.14.718463

Figure Lengend Snippet: (a) CD31 expression in HCAEC treated with TGFβ1 (10ng/ml), Hypoxia (5% O 2 ), or TGFβ1+ Hypoxia in the absence or presence of moderate dose ethanol (EtOH 25 mM). (b) αSMA expression in HCAEC treated with TGFβ +/- either moderate dose ethanol (25 mM EtOH) or high dose ethanol (100 mM EtOH). (c) SM22α and (d) αSMA expression in HUVEC treated with IL1β+TGFβ2 in the absence or presence of either 25 mM EtOH or 100 mM EtOH as indicated. Representative immunofluorescence images, from at least 3 experiments, shown.

Article Snippet: Primary antibodies used include human CD31 (Cat. # AF806, R&D Systems), αSMA (MAB1420, R&D systems), SM22α (Cat. # 36090 CST), and CDH5 (Cat. # 2500, CST).

Techniques: Expressing, Immunofluorescence

Journal: bioRxiv

Article Title: A Biphasic Effect of Alcohol on Endothelial Plasticity Through Regulation of Endothelial-to-Mesenchymal Transition

doi: 10.64898/2026.04.14.718463

Figure Lengend Snippet: (a) (b) Representative images of immunofluorescently stained carotid cross sections from sham-operated and ligated controls, moderate EtOH, and Binge EtOH experimental groups (males). Blue = Dapi nuclear stain, red = Tm-Cdh5+, and white=αSMA+. (c) Cells co-expressing Cdh5 and αSMA (i.e., myo-endothelial cells indicative of EndMT) were quantified using QuPath bioimage analysis software in carotid cross sections post-ligation from controls (grey bars), moderate EtOH (green bars), and binge EtOH (red bars) experimental groups. Bar graphs show cumulative data expressed as % Myoendothelial cells /full cross section, % Myoendothelial cells /neointima, or Number of myoendothelial cells/full cross section. Data are mean±SEM, n=25-30 sections from 5-6 mice). *p<0.05, ** P<0.001, **** p<0.0001.

Article Snippet: Primary antibodies used include human CD31 (Cat. # AF806, R&D Systems), αSMA (MAB1420, R&D systems), SM22α (Cat. # 36090 CST), and CDH5 (Cat. # 2500, CST).

Techniques: Staining, Expressing, Software, Ligation

Journal: bioRxiv

Article Title: Monocytes Strongly Induce (MYO)Fibroblast Contraction in a New 3D Skin Model to Understand the Inflammation-Fibrosis Axis in Systemic Sclerosis

doi: 10.64898/2026.02.12.705496

Figure Lengend Snippet: ( A ) Sorted CD45-cells (fibroblasts) were analyzed for expression of myofibroblast-related genes after co-culture with monocytes in collagen hydrogels. Values (mean ± SEM) indicate relative gene expression (−ΔCt) measured with qPCR. Reference genes used were GAPDH and RPS27A. Analyses were done by two-way ANOVA with Sidak’s multiple comparisons test (**p<0.01, ***p<0.001, ****p<0.0001). ( B ) Representative immunohistochemistry staining showing robust expression of α-SMA (alpha-smooth muscle actin) (left) and fibroblast activation protein (FAP) (right) in the hydrogels containing only fibroblasts (top) compared to those co-culture with monocytes (bottom). The small blue nuclei in the bottom panels are monocytes. (Myo)fibroblasts are clearly positive for both markers (black arrows), with a typically elongated shape.

Article Snippet: Sections were then incubated with primary rabbit –anti-human alpha-smooth muscle actin (α-SMA) (1:200) (ab5694, Abcam, UK), – fibroblast activation protein (FAP) (1:100) (ab207178, Abcam), – Phospho-SMAD2 (1:100) (#3108L, CellSignaling, USA), – Phospho-Stat3 (1:50) (#9145, CellSignaling), or mouse anti-human – CD68 (#MCA1815, BioRad, USA) (1:200) antibodies, for 1 hour at room temperature.

Techniques: Expressing, Co-Culture Assay, Gene Expression, Immunohistochemistry, Staining, Activation Assay

Journal: bioRxiv

Article Title: Monocytes Strongly Induce (MYO)Fibroblast Contraction in a New 3D Skin Model to Understand the Inflammation-Fibrosis Axis in Systemic Sclerosis

doi: 10.64898/2026.02.12.705496

Figure Lengend Snippet: ( A ) Sorted CD45-cells (fibroblasts) were analyzed for expression of myofibroblast-related genes after co-culture with monocytes in collagen hydrogels. Values (mean ± SEM) indicate relative gene expression (−ΔCt) measured with qPCR. Reference genes used were GAPDH and RPS27A. Analyses were done by two-way ANOVA with Sidak’s multiple comparisons test (**p<0.01, ***p<0.001, ****p<0.0001). ( B ) Representative immunohistochemistry staining showing robust expression of α-SMA (alpha-smooth muscle actin) (left) and fibroblast activation protein (FAP) (right) in the hydrogels containing only fibroblasts (top) compared to those co-culture with monocytes (bottom). The small blue nuclei in the bottom panels are monocytes. (Myo)fibroblasts are clearly positive for both markers (black arrows), with a typically elongated shape.

Article Snippet: Sections were then incubated with primary rabbit –anti-human alpha-smooth muscle actin (α-SMA) (1:200) (ab5694, Abcam, UK), – fibroblast activation protein (FAP) (1:100) (ab207178, Abcam), – Phospho-SMAD2 (1:100) (#3108L, CellSignaling, USA), – Phospho-Stat3 (1:50) (#9145, CellSignaling), or mouse anti-human – CD68 (#MCA1815, BioRad, USA) (1:200) antibodies, for 1 hour at room temperature.

Techniques: Expressing, Co-Culture Assay, Gene Expression, Immunohistochemistry, Staining, Activation Assay