Journal: PLoS Neglected Tropical Diseases

Article Title: A Polarized Cell Model for Chikungunya Virus Infection: Entry and Egress of Virus Occurs at the Apical Domain of Polarized Cells

doi: 10.1371/journal.pntd.0002661

Figure Lengend Snippet: ( A ) CHIKV-infected HBMEC cells were viewed under DIC microscopy to observe for morphological changes. The cell density was lower in CHIKV-infected cells as compared to mock-infected cells from 48 h.p.i. onwards. ( B ) CHIKV-infected HBMEC cells were fixed at various intervals post-infection and immunofluorescence assay was performed to detect for CHIKV protein expression (green). Cell nuclei were stained with DAPI (blue). Moderate amounts of CHIKV protein expression were observed at 24 h.p.i. of HBMEC. ( C ) Quantification of infectious virus titer by plaque assay showed an increasing trend, with a peak in infectious virus titer at 36 h.p.i.. Vertical bars represent one standard deviation from the mean of three readings.

Article Snippet: Human Brain Microvascular Endothelial Cells (HBMEC) from ScienCell were maintained in Endothelial Cell Medium (ECM) supplemented with 5% fetal bovine serum (FBS) and 1% endothelial cell growth supplement.

Techniques: Infection, Microscopy, Immunofluorescence, Expressing, Staining, Virus, Plaque Assay, Standard Deviation

Journal: PLoS Neglected Tropical Diseases

Article Title: A Polarized Cell Model for Chikungunya Virus Infection: Entry and Egress of Virus Occurs at the Apical Domain of Polarized Cells

doi: 10.1371/journal.pntd.0002661

Figure Lengend Snippet: ( A ) TEER measurements of Vero, Vero C1008 and HBMEC cell monolayers were taken at 24 h.p.i.. The TEER measurements post-apical and basolateral infection were comparable to that of mock-infected cells. Vertical bars represent one standard deviation from the mean of three readings. ( B ) Immunofluorescence assays demonstrated the expression of ZO-1 tight junction proteins (green) in apically-infected and basolaterally-infected Vero, Vero C1008 and HBMEC cells at 24 h.p.i.. The expression of ZO-1 proteins in infected cells is comparable to that of mock-infected cells. (C) The FITC-dextran permeability assays demonstrated that the integrity of Vero and Vero C1008 cell monolayers remained intact at 24 h.p.i., where the permeability of the cell monolayers to FITC-dextran remained low as compared to the TNF-treated cells.

Article Snippet: Human Brain Microvascular Endothelial Cells (HBMEC) from ScienCell were maintained in Endothelial Cell Medium (ECM) supplemented with 5% fetal bovine serum (FBS) and 1% endothelial cell growth supplement.

Techniques: Infection, Standard Deviation, Immunofluorescence, Expressing, Permeability

Journal: PLoS Neglected Tropical Diseases

Article Title: A Polarized Cell Model for Chikungunya Virus Infection: Entry and Egress of Virus Occurs at the Apical Domain of Polarized Cells

doi: 10.1371/journal.pntd.0002661

Figure Lengend Snippet: ( A ) The total virus yield at 24 h.p.i. of non-polarized Vero, polarized Vero C1008 and polarized HBMEC cells at an MOI of 10 was quantified by viral plaque assay. Entry of CHIKV is bi-directional in non-polarized Vero cells but occurs preferentially at the apical domain of polarized Vero C1008 and HBMEC cells. Two-tailed Student's t -test: * p<0.05, ** p<0.005. Vertical bars represent one standard deviation from the mean of three readings. ( B ) Virus binding assays show higher amount of CHIKV binding upon apical infection as compared to upon basolateral infection of polarized Vero C1008 cells. In contrast, the amount of CHIKV binding upon apical and basolateral infection of non-polarized Vero cells is similar. Two-tailed Student's t -test: *** p<0.001.

Article Snippet: Human Brain Microvascular Endothelial Cells (HBMEC) from ScienCell were maintained in Endothelial Cell Medium (ECM) supplemented with 5% fetal bovine serum (FBS) and 1% endothelial cell growth supplement.

Techniques: Virus, Viral Plaque Assay, Two Tailed Test, Standard Deviation, Binding Assay, Infection

Journal: PLoS Neglected Tropical Diseases

Article Title: A Polarized Cell Model for Chikungunya Virus Infection: Entry and Egress of Virus Occurs at the Apical Domain of Polarized Cells

doi: 10.1371/journal.pntd.0002661

Figure Lengend Snippet: ( A ) Infectious virus titer of supernatants collected from the apical and basolateral chambers at 24 h.p.i. of non-polarized Vero, polarized Vero C1008 and polarized HBMEC cells at an MOI of 10 were quantified by viral plaque assays. Release of CHIKV is bi-directional in non-polarized Vero cells but occurs preferentially at the apical domain of polarized Vero C1008 and HBMEC cells. Two-tailed Student's t -test: * p<0.05, ** p<0.005, *** p<0.001. Vertical bars represent one standard deviation from the mean of three readings. ( B ) Apically-infected Vero C1008, ( C ) basolaterally-infected Vero C1008, ( D ) apically-infected Vero and ( E ) basolaterally-infected Vero cells were co-labeled with antibodies against CHIKV E2 glycoprotein (green, arrows) and ZO-1 (red, arrowheads). Cell nuclei were stained with DAPI (blue). Z-stacked images show the polarized release of CHIKV towards the apical plasma membrane of Vero C1008 upon apical and basolateral infection. Release of CHIKV is bidirectional in non-polarized Vero cells.

Article Snippet: Human Brain Microvascular Endothelial Cells (HBMEC) from ScienCell were maintained in Endothelial Cell Medium (ECM) supplemented with 5% fetal bovine serum (FBS) and 1% endothelial cell growth supplement.

Techniques: Virus, Two Tailed Test, Standard Deviation, Infection, Labeling, Staining, Clinical Proteomics, Membrane

Journal:

Article Title: Anti-Tumoral Activity of a Short Decapeptide Fragment of the Alzheimer's A? Peptide

doi: 10.1007/s10989-010-9198-8

Figure Lengend Snippet: Effect of 4-hydroxy tamoxifen on angiogenesis in vitro. a Representative pictures depicting the formation of capillary networks in response to a dose range of 4-hydroxy tamoxifen following the plating of human brain microvascular endothelial cells onto a layer of Matrigel. b The graph represents the quantification of capillary network length by image analysis and the amount of LDH activity detected in the culture medium surrounding the capillary like structures following 24 h of treatment with a dose range of 4-hydroxy tamoxifen. ANOVA revealed a significant main effect of 4-hydroxy tamoxifen on LDH released (P < 0.03) but no significant main effect on capillary formation (P = 0.743) and post hoc analysis showed significant differences for LDH released between control conditions and 10 µM of 4-hydroxy tamoxifen (P < 0.009)

Article Snippet: Capillary Morphogenesis Assay Human Brain Microvascular Endothelial Cells (HBMEC) (Sciencell, CA) were cultured in Endothelial Cell Growth Medium (Cell Applications, CA) containing 5% fetal bovine serum, 1% penicillin/streptomycin and 1% Endothelial Cell Growth Supplement (Sigma–Aldrich, MO).

Techniques: In Vitro, Activity Assay, Control