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(A) The binding between purified prothrombin and V5-His-RPN2 (filled circle) was confirmed by ELISA. BSA (filled triangle) was used as the control. The data shown in this figure were from one representative experiment of three experiments. (B) The binding between purified prothrombin as ligand and V5-His-RPN2 (1.00 µM; solid line) as analyte was investigated by surface plasmon resonance. BSA (dashed line) was used as a control. Response rate is 200 resonance units (RU). RPN2: ribophorin II. From: Ribophorin II is involved in the tissue factor expression mediated by phosphatidylserine-dependent antiprothrombin antibody on monocytes Rheumatology (Oxford). 2016;55(6): doi: /rheumatology/kew005 Rheumatology (Oxford) | © The Author Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For Permissions, please
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(A) Expression of secretory prothrombin
(A) Expression of secretory prothrombin. HEK293 cells were transfected with expression plasmids encoding FLAG-tagged prothrombin. Cells were lysed and subjected to immunoblot analysis with anti-FLAG or anti-prothrombin antibody. (B) Pull-down analysis of proteins binding to FLAG-prothrombin. Coomasie brilliant green staining of prothrombin-associated proteins. From: Ribophorin II is involved in the tissue factor expression mediated by phosphatidylserine-dependent antiprothrombin antibody on monocytes Rheumatology (Oxford). 2016;55(6): doi: /rheumatology/kew005 Rheumatology (Oxford) | © The Author Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For Permissions, please
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(A) The efficacy of RPN2-siRNA in RAW264
(A) The efficacy of RPN2-siRNA in RAW264.7 cells was assessed 48 h after transfection by quantitative real-time PCR (qPCR). The expression of RPN2 mRNA in RPN2-siRNA-treated cells is grey, control siRNA is black and non-treatment is hatched. The bars represent the mean (s.e.m.) of eight independent experiments. (B) The efficacy of RPN2-siRNA was assessed 72 h after transfection by flow cytometry. The expression level of RPN2 on the cell surface in siRNA-treated cells is white and control siRNA is grey. *P < RPN2: ribophorin II; siRNA: small interfering RNA. From: Ribophorin II is involved in the tissue factor expression mediated by phosphatidylserine-dependent antiprothrombin antibody on monocytes Rheumatology (Oxford). 2016;55(6): doi: /rheumatology/kew005 Rheumatology (Oxford) | © The Author Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For Permissions, please
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(A) Intracellular expression of RPN2 was investigated with permeabilization solution. The expression level of intracellular RPN2 is white and control IgG is grey. (B) Cell surface expression of RPN2 was confirmed and investigated without permeabilization solution. The expression level of RPN2 on the cell surface is white and control IgG is grey. RPN2: ribophorin II. From: Ribophorin II is involved in the tissue factor expression mediated by phosphatidylserine-dependent antiprothrombin antibody on monocytes Rheumatology (Oxford). 2016;55(6): doi: /rheumatology/kew005 Rheumatology (Oxford) | © The Author Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For Permissions, please
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Prothrombin, monoclonal aPS-PT (231D) and mouse IgG (control IgG) were added to RAW264.7 cells treated with RPN2-siRNA (grey) or control-siRNA (black) at a concentration of 10 µg/ml. Lipopolysaccharide was used at a concentration of 200 ng/ml. The cells were treated for 4 h to determine TF mRNA. *P< RPN2: ribophorin II; siRNA: small interfering RNA; TF, tissue factor. From: Ribophorin II is involved in the tissue factor expression mediated by phosphatidylserine-dependent antiprothrombin antibody on monocytes Rheumatology (Oxford). 2016;55(6): doi: /rheumatology/kew005 Rheumatology (Oxford) | © The Author Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For Permissions, please
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