human brain microvascular endothelial cells (hbvecs) sciencell Search Results


human brain microvascular endothelial cells (hbvecs) sciencell #1000  (ScienCell)
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ScienCell human brain microvascular endothelial cells (hbvecs) sciencell #1000
a ) 20 PFU of WNV was inoculated intracranially. At day 5 p.i., brain viral loads (shown by WNV-E expression) and cytokine/chemokine or chemokine receptor transcripts were quantified by q-PCR and normalized with mouse actin. b–d, f ) Mice were infected with 200 PFU of WNV via s.c. footpad injection. b ) Expression of select chemokine receptors by blood cells was quantified by q-PCR using specific primers and probes, and normalized with beta actin. c ) The Cxcr2 expression levels of whole blood cells at day 4 p.i. were determined by immunostaining and flow cytometry. The results are expressed as mean fluorescence intensity (MFI) per cell. d ) The mRNA levels of cxcl1 and cxcl5 in WT and Il22 −/− brains were assessed by q-PCR. Each dot in the plot represents one mouse; * p <0.05; ** p <0.01. e ) Expression of CXCL1 and CXCL5 mRNA by human brain <t>microvascular</t> <t>endothelial</t> cells (HBVEC) treated with human recombinant IL-22. Bars: the mean of the results with standard deviation. ** p <0.01 ( n = 3). f ) Numbers of human PMNs that migrated out of the upper chamber (2×10 5 ) across the HBVEC monolayer down to the lower chamber over 18 hours in the presence of various concentrations of IL-22. Each dot represents the mean cell number from one well (counted 4 times). ** p <0.001. g ) Numbers of mouse PMNs that migrated out of the upper chamber (2×10 5 ) across the mouse brain vascular endothelial cell (MBVEC) monolayer down to the lower chamber. Each dot represents the mean cell number from one well (counted 4 times). * p <0.05. h ) Quantification of Cxcl1 levels of MBVECs treated as in g ) by ELISA. i ) Immunofluorescence staining of WNV-E (green), neutrophils (7/4, red) and DAPI (blue) in the brain cortex sections at day 5 p.i. Arrows point to co-staining. Micrographs were acquired using a laser scanning confocal microscope (20X images are shown and are representative of n = 5 mice per group). A higher magnification (63x) of confocal images of WT brain section is shown below (WNV, red; 7/4, green, TO-PRO3, blue). Arrow heads indicate typical polynuclear morphology of neutrophils. j ) FACS analyses of brain neutrophils (CD45Ly6G + ) of WT and Il22 −/− mice on day 6 p.i. Each dot represents 2 mice. * p <0.05.
Human Brain Microvascular Endothelial Cells (Hbvecs) Sciencell #1000, supplied by ScienCell, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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a ) 20 PFU of WNV was inoculated intracranially. At day 5 p.i., brain viral loads (shown by WNV-E expression) and cytokine/chemokine or chemokine receptor transcripts were quantified by q-PCR and normalized with mouse actin. b–d, f ) Mice were infected with 200 PFU of WNV via s.c. footpad injection. b ) Expression of select chemokine receptors by blood cells was quantified by q-PCR using specific primers and probes, and normalized with beta actin. c ) The Cxcr2 expression levels of whole blood cells at day 4 p.i. were determined by immunostaining and flow cytometry. The results are expressed as mean fluorescence intensity (MFI) per cell. d ) The mRNA levels of cxcl1 and cxcl5 in WT and Il22 −/− brains were assessed by q-PCR. Each dot in the plot represents one mouse; * p <0.05; ** p <0.01. e ) Expression of CXCL1 and CXCL5 mRNA by human brain microvascular endothelial cells (HBVEC) treated with human recombinant IL-22. Bars: the mean of the results with standard deviation. ** p <0.01 ( n = 3). f ) Numbers of human PMNs that migrated out of the upper chamber (2×10 5 ) across the HBVEC monolayer down to the lower chamber over 18 hours in the presence of various concentrations of IL-22. Each dot represents the mean cell number from one well (counted 4 times). ** p <0.001. g ) Numbers of mouse PMNs that migrated out of the upper chamber (2×10 5 ) across the mouse brain vascular endothelial cell (MBVEC) monolayer down to the lower chamber. Each dot represents the mean cell number from one well (counted 4 times). * p <0.05. h ) Quantification of Cxcl1 levels of MBVECs treated as in g ) by ELISA. i ) Immunofluorescence staining of WNV-E (green), neutrophils (7/4, red) and DAPI (blue) in the brain cortex sections at day 5 p.i. Arrows point to co-staining. Micrographs were acquired using a laser scanning confocal microscope (20X images are shown and are representative of n = 5 mice per group). A higher magnification (63x) of confocal images of WT brain section is shown below (WNV, red; 7/4, green, TO-PRO3, blue). Arrow heads indicate typical polynuclear morphology of neutrophils. j ) FACS analyses of brain neutrophils (CD45Ly6G + ) of WT and Il22 −/− mice on day 6 p.i. Each dot represents 2 mice. * p <0.05.

Journal: PLoS ONE

Article Title: IL-22 Signaling Contributes to West Nile Encephalitis Pathogenesis

doi: 10.1371/journal.pone.0044153

Figure Lengend Snippet: a ) 20 PFU of WNV was inoculated intracranially. At day 5 p.i., brain viral loads (shown by WNV-E expression) and cytokine/chemokine or chemokine receptor transcripts were quantified by q-PCR and normalized with mouse actin. b–d, f ) Mice were infected with 200 PFU of WNV via s.c. footpad injection. b ) Expression of select chemokine receptors by blood cells was quantified by q-PCR using specific primers and probes, and normalized with beta actin. c ) The Cxcr2 expression levels of whole blood cells at day 4 p.i. were determined by immunostaining and flow cytometry. The results are expressed as mean fluorescence intensity (MFI) per cell. d ) The mRNA levels of cxcl1 and cxcl5 in WT and Il22 −/− brains were assessed by q-PCR. Each dot in the plot represents one mouse; * p <0.05; ** p <0.01. e ) Expression of CXCL1 and CXCL5 mRNA by human brain microvascular endothelial cells (HBVEC) treated with human recombinant IL-22. Bars: the mean of the results with standard deviation. ** p <0.01 ( n = 3). f ) Numbers of human PMNs that migrated out of the upper chamber (2×10 5 ) across the HBVEC monolayer down to the lower chamber over 18 hours in the presence of various concentrations of IL-22. Each dot represents the mean cell number from one well (counted 4 times). ** p <0.001. g ) Numbers of mouse PMNs that migrated out of the upper chamber (2×10 5 ) across the mouse brain vascular endothelial cell (MBVEC) monolayer down to the lower chamber. Each dot represents the mean cell number from one well (counted 4 times). * p <0.05. h ) Quantification of Cxcl1 levels of MBVECs treated as in g ) by ELISA. i ) Immunofluorescence staining of WNV-E (green), neutrophils (7/4, red) and DAPI (blue) in the brain cortex sections at day 5 p.i. Arrows point to co-staining. Micrographs were acquired using a laser scanning confocal microscope (20X images are shown and are representative of n = 5 mice per group). A higher magnification (63x) of confocal images of WT brain section is shown below (WNV, red; 7/4, green, TO-PRO3, blue). Arrow heads indicate typical polynuclear morphology of neutrophils. j ) FACS analyses of brain neutrophils (CD45Ly6G + ) of WT and Il22 −/− mice on day 6 p.i. Each dot represents 2 mice. * p <0.05.

Article Snippet: Primary human brain microvascular endothelial cells (HBVECs) (ScienCell, #1000) were cultured in formulated ECM medium (ScienCell, #1001) and treated with human recombinant IL-22 at a concentration of 200 ng/ml.

Techniques: Expressing, Infection, Injection, Immunostaining, Flow Cytometry, Fluorescence, Recombinant, Standard Deviation, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Microscopy