Journal: Scientific Reports

Article Title: Endothelial glycocalyx shields the interaction of SARS-CoV-2 spike protein with ACE2 receptors

doi: 10.1038/s41598-021-91231-1

Figure Lengend Snippet: Differences in adhesive interactions between the S-proteins of SARS-CoV-2 and the surfaces of human bronchial epithelial cells (HBECs) and human pulmonary artery endothelial cells (HPAECs). ( a ) Histograms of the maximal detachment force F max for HBECs. Left: comparison of data for native and anti-ACE2 treated cells. Right: comparison of data for native and heparin treated system. ( b ) Comparison of mean values determined from histograms. ( c ) AFM height map measured for a single HBEC cell. ( d ) Corresponding adhesive maps measured for this cell. Left: native cell. Middle: anti-ACE2 treatment. Right: heparin treatment. ( e ) Fluorescent staining of ACE2 (green, left column), glycocalyx (Glx, red, middle column) and Merged (right column). ( f ) Quantitative data of ACE2 and Glx mean fluorescence intensity. ( g ) Histograms of the maximal detachment force F max for HPAECs. Left: comparison of data for native and anti-ACE2 treated cells. Right: comparison of data for native and heparin treated system. ( h ) Comparison of mean values determined from histograms. ( i ) AFM height map measured for a single HPAEC cell. ( j ) Corresponding adhesive maps measured for this cell. ( k ) Fluorescent staining of ACE2 (green, left column), Glx (red, middle column) and Merged (right column) ( l ) Quantitative data of ACE2 and Glx mean fluorescence intensity. Left: native cell. Middle: anti-ACE2 treatment. Right: heparin treatment. Statistics: p values were determined by one-way ANOVA followed by Tukey’s post-hoc test. Experimental details are listed in Supplementary Table . Source data are provided as a Source Data file. Figure created with OriginPro2021 ( https://www.originlab.com/2021 ), ImageJ 1.53e ( https://imagej.nih.gov/ij/ ) and JPK Data Processing 6.1.79 ( https://www.jpk.com/ ).

Article Snippet: Primary Human Pulmonary Artery Endothelial Cells (HPAECs, ATCC) were grown in Vascular Cell Basal Medium (Cat. No. PCS-100-030, ATCC), supplemented with Endothelial Cell Growth Kit-VEGF (Cat. No. PCS-100-041, ATCC) The cells were maintained in standard conditions at 37 °C, 5% CO 2 , and 95% humidity.

Techniques: Adhesive, Comparison, Staining, Fluorescence

Journal: Scientific Reports

Article Title: Endothelial glycocalyx shields the interaction of SARS-CoV-2 spike protein with ACE2 receptors

doi: 10.1038/s41598-021-91231-1

Figure Lengend Snippet: Endothelial cells stiffening after incubation with S-protein is more pronounced for cells with removed glycocalyx. ( a ) Elastic modulus of HPAECs for native cells (grey histogram) and for cells incubated with S-protein (magenta). ( b ) Mean fluorescence intensity of phalloidin (AlexaFluor488). ( c ) Examples of fluorescence images depict the actin structure in native HPAECs and after incubation with S-protein. Green—actin. Blue—nucleus. ( d ) Examples of AFM-QI images of native HPAECs and after incubation with S-protein. ( e ) Elastic modulus obtained for HPAECs pre-incubated with heparinase (green) and next incubated with S-protein (magenta). ( f ) Mean fluorescence intensity of phalloidin (AlexaFluor488) after removal of HS. ( g ) Fluorescence images show the actin polymerization that occurred after incubation with S-protein for HPAECs pre-incubated with heparinase. ( h ) Examples of AFM-QI images depict the changes of cell morphology and cortical actin network after incubation with S-protein for HPAECs pre-incubated with heparinase. Statistics: p values were determined by one-way ANOVA followed by Tukey’s post-hoc test. Figure created with OriginPro2021 ( https://www.originlab.com/2021 ), ImageJ 1.53e ( https://imagej.nih.gov/ij/ ) and JPK Data Processing 6.1.79 ( https://www.jpk.com/ ).

Article Snippet: Primary Human Pulmonary Artery Endothelial Cells (HPAECs, ATCC) were grown in Vascular Cell Basal Medium (Cat. No. PCS-100-030, ATCC), supplemented with Endothelial Cell Growth Kit-VEGF (Cat. No. PCS-100-041, ATCC) The cells were maintained in standard conditions at 37 °C, 5% CO 2 , and 95% humidity.

Techniques: Incubation, Fluorescence

Journal: Toxins

Article Title: Isolation and Biochemical Characterization of Rubelase, a Non-Hemorrhagic Elastase from Crotalus ruber ruber (Red Rattlesnake) Venom

doi: 10.3390/toxins3070900

Figure Lengend Snippet: Cytotoxic effects of rubelase and rubelysin on cultured cells. ( A ) HUVEC: human umbilical vein endothelial cells; ( B ) HPAEC: human pulmonary artery endothelial cells; ( C ) HASMC: human aortic smooth muscle cells. Rubelase and rubelysin were added to the cells at various concentrations. After incubation for 18 h, viable cells were counted using the colorimetric method. The results shown represent the average of five experiments. The absorbance of cultured cells incubated with saline or crude venom (10 µg) was defined as control and (cytotoxic) positive control *, respectively.

Article Snippet: Cryo-preserved human umbilical vein endothelial cells (HUVEC), human pulmonary artery endothelial cells (HPAEC), human aortic smooth muscle cells (HASMC), and their respective cell culture medium were obtained from Kurabo (Japan).

Techniques: Cell Culture, Incubation, Saline, Control, Positive Control

Journal: Toxins

Article Title: Isolation and Biochemical Characterization of Rubelase, a Non-Hemorrhagic Elastase from Crotalus ruber ruber (Red Rattlesnake) Venom

doi: 10.3390/toxins3070900

Figure Lengend Snippet: Fluorescence micrographs of HPAEC (×60) after incubation with rubelase and rubelysin. Control cells ( A ); and the cells incubated with rubelase ( B ); and rubelysin ( C ).

Article Snippet: Cryo-preserved human umbilical vein endothelial cells (HUVEC), human pulmonary artery endothelial cells (HPAEC), human aortic smooth muscle cells (HASMC), and their respective cell culture medium were obtained from Kurabo (Japan).

Techniques: Fluorescence, Incubation, Control