Cultured human glomerular mesangial cells express the C5a receptor

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Cultured human glomerular mesangial cells express the C5a receptor Michael Braun, Alvin E. Davis Kidney International Volume 54, Issue 5, Pages 1542-1549 (November 1998) DOI: 10.1046/j.1523-1755.1998.00155.x Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 1 Expression of the C5a receptor by cultured human glomerular mesangial cells (HGMC). (A) Cell surface immunofluorescence of HGMC using rabbit anti-C5a receptor antibody. (B) Negative control using rabbit anti-hemoglobin antibody. Magnification is ×40. Kidney International 1998 54, 1542-1549DOI: (10.1046/j.1523-1755.1998.00155.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 2 FACS analysis of cultured human glomerular mesangial cells (HGMC) using an FITC-labeled murine monoclonal antibody to the C5a receptor in comparison with FITC-labeled isotype matched control antibody. Kidney International 1998 54, 1542-1549DOI: (10.1046/j.1523-1755.1998.00155.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 3 Reverse transcriptase-polymerase chain reaction (RT-PCR) identification of C5a receptor mRNA in human glomerular mesangial cells (HGMC). Samples were subjected to electrophoresis on a 5% polyacrylamide gel and stained with ethidium bromide. Lane 1, 25bp ladder; lane 2, blank; lane 3, blank; lane 4, RT-PCR amplified mRNA from U937 cells; lane 5, RT-PCR amplified mRNA from human liver; lane 6, RT-PCR amplified mRNA from HGMC. In both the U937 cells and the HGMC, only the mRNA specific 209bp product was observed. Kidney International 1998 54, 1542-1549DOI: (10.1046/j.1523-1755.1998.00155.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 4 C5a-mediated enhanced proliferation of human glomerular mesangial cells (HGMC). Quadruplicate HGMC samples were either untreated or treated with 10 nm, 50 nm, 100 nm and 200 nm rC5a. Neutral red dye uptake was quantitated at 24, 48 and 72hours, as described in the Methods section. Symbols are: (•) control; (○) 10 nm; (▾) 50 nm; (▿) 100 nm; (▪) 200 nm. Kidney International 1998 54, 1542-1549DOI: (10.1046/j.1523-1755.1998.00155.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 5 Induction of monocyte chemoattractant protein-1 (MCP-1) in human glomerular mesangial cell (HGMC) supernatants. (A) rC5a dose-response at 24hours post-stimulation. (B) Time course analysis of MCP-1 expression following stimulation with 25 nm rC5a. Each bar plots the mean of triplicate samples with the bracket indicating 1 sd. Symbols are: (∗) control; (#) 10 nm; (†) 25 nm; (‡) 50 nm; (§) 100 nm.**P = 0.01. Kidney International 1998 54, 1542-1549DOI: (10.1046/j.1523-1755.1998.00155.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 6 Induction of platelet-derived growth factor-AB (PDGF-AB) in human glomerular mesangial cell (HGMC) supernatants. (A) rC5a dose-response at 24hours post-stimulation. (B) Time course analysis of PDGF-AB expression following stimulation with 25 nm rC5a. Each bar plots the mean of triplicate samples with the bracket indicating 1 sd. Symbols are: (∗) control; (#) 10 nm; (†) 25 nm; (‡) 50 nm; (§) 100 nm.**P = 0.003. Kidney International 1998 54, 1542-1549DOI: (10.1046/j.1523-1755.1998.00155.x) Copyright © 1998 International Society of Nephrology Terms and Conditions