Journal: bioRxiv

Article Title: Direct effects on endothelial cells are essential for perivascular cell (pericyte)-dependent amplification of orthoflavivirus NS1-mediated microvascular leakage

doi: 10.64898/2026.01.29.702573

Figure Lengend Snippet: (A-B) ECIS measurement of permeability of monocultures of HUVECs (A) or LSECs (B) treated with preconditioned media from HSCs treated with 1 µg/mL NS1 or 100 ng/mL TNF-α for 24 h; “Untreated” refers to HUVECs or LSECs treated with HSC media only and the “EC only” refers to untreated HUVECs or LSECs without the addition of HSC media. (C, D) Dextran permeability assay measuring HUVEC (C) or LSEC (D) permeability in coculture with HSCs 24 h post-treatment with 1 µg/mL NS1, 100 ng/mL TNF-α (positive control), or eluate (negative) control. Data points represent technical replicates within an experiment with different symbol shapes corresponding to different experiments (N=3, n=3). Bars represent the mean, and error bars represent SEM. (E, F) TEER measurement of semi-contact cocultures of HUVECs (A) or LSECs (B) cultured with HSCs and treated with 1 µg/mL NS1, 100 ng/mL TNF-α (positive control), or eluate (negative) control. Permeability measurements were normalised to each sample pre-treatment for ECIS data (A, B) or the untreated endothelial cell only control (EC only; dashed line) for EVOM data (E, F). In the time course experiments (A, B, E, F), each data point represents the mean ± SEM. In all experiments indicated significance is compared to untreated cocultures; * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were obtained from PromoCell (Heidelberg, Germany); liver sinusoidal endothelial cells (LSECs) from Innoprot (Derio, Spain), and human hepatic stellate cells (HSCs) from Zen-Bio (Durham, NC USA).

Techniques: Permeability, FITC-Dextran Permeability Assay, Positive Control, Negative Control, Cell Culture, Control

Journal: bioRxiv

Article Title: Direct effects on endothelial cells are essential for perivascular cell (pericyte)-dependent amplification of orthoflavivirus NS1-mediated microvascular leakage

doi: 10.64898/2026.01.29.702573

Figure Lengend Snippet: Endothelial barrier integrity measurements for HUVEC (A, B) or LSEC (C) monocultures following treatment with 1 µg/mL wild-type DENV-2, AHFV or KFDV NS1, or the nonfunctional DENV-2 NS207Q mutant (negative control), or 100 ng/mL TNF-α (positive control), or eluate (negative) control as indicated. Permeability measurements were normalised to the untreated endothelial cell only control (Untreated) for EVOM TEER data (A), or to each sample before treatment for ECIS data (B, C). Each data point represents the mean ± SEM. Indicated significance is for treatment versus untreated EC only control (Untreated). * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were obtained from PromoCell (Heidelberg, Germany); liver sinusoidal endothelial cells (LSECs) from Innoprot (Derio, Spain), and human hepatic stellate cells (HSCs) from Zen-Bio (Durham, NC USA).

Techniques: Mutagenesis, Negative Control, Positive Control, Permeability, Control

Journal: Journal of Leukocyte Biology

Article Title: Frontline Science: HIV infection of Kupffer cells results in an amplified proinflammatory response to LPS

doi: 10.1189/jlb.3HI0516-242R

Figure Lengend Snippet: (A) Immunofluorescence CD68 staining (green) reveals KCs within the hepatic sinusoids (left, 200× original magnification; right, 640× original magnification). (B) Number of KCs/mm2 quantitated and shown graphically. HCV and HIV/HCV, n = 6; normal, n = 3. (C) KC morphology (phase-contrast photomicrograph original magnification, 200×). (D) Representative FACS analyses of CD68 expression directly after KC isolation (gray histogram) and adherence enrichment of KCs (white histogram). (E) FACS demonstrating quantitative expression of CD68 in primary liver-derived KCs directly after isolation, with adherence enrichment, and after HIV infection (n = 10). (F) RT-qPCR confirms low expression of CD3 and CD31 in KC preparations compared with human T cells and liver sinusoidal endothelial cells (LSECs), respectively (n = 3 T cells and LSECs; n = 5 KCs).

Article Snippet: RT-qPCR on RNA extracted from KCs was performed to determine the presence of significant contamination with T cells (primers to CD3) or endothelial cells (primers to CD31). cDNA from primary CD4 + T cells and human hepatic sinusoidal endothelial cells (Cat. #5000; ScienCell, Carlsbad, CA, USA) were used as positive controls for CD3 and CD31 expression, respectively.

Techniques: Immunofluorescence, Staining, Expressing, Isolation, Derivative Assay, Infection, Quantitative RT-PCR