Lipoprotein-stimulated mesangial cell proliferation and gene expression are regulated by lipoprotein lipase1  Frazier T. Stevenson, Gregory C. Shearer,

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Lipoprotein-stimulated mesangial cell proliferation and gene expression are regulated by lipoprotein lipase1  Frazier T. Stevenson, Gregory C. Shearer, David N. Atkinson  Kidney International  Volume 59, Issue 6, Pages 2062-2068 (June 2001) DOI: 10.1046/j.1523-1755.2001.00720.x Copyright © 2001 International Society of Nephrology Terms and Conditions

Figure 1 Binding of rat very low-density lipoprotein (VLDL) to primary rat mesangial cells (MCs) at 4°C. Abbreviations are: H'ase, 1 IU/mL heparanase pretreatment of cells for one hour prior to binding study; LpL, treatment of cells with 3 μg/mL bovine milk LpL for four hours prior to binding study. N = 3 to 4 for each VLDL concentration used. Symbols are: (▪) VLDL + LpL; (▾) VLDL + LpL + heparanase; (♦) VLDL + heparanase; (▴) VLDL; *P < 0.001 vs. all other groups. Kidney International 2001 59, 2062-2068DOI: (10.1046/j.1523-1755.2001.00720.x) Copyright © 2001 International Society of Nephrology Terms and Conditions

Figure 2 Mesangial cell (MC) proliferation in response to very low-density lipoprotein (VLDL) and lipoprotein lipase (LpL). All cells were quiesced for 24 hours prior to stimulation for 24 hours. Proliferation is expressed as increase in Quantose dye fluorescence compared with fluorescence after quiescence. N = 5 to 7 per group. Symbols are: (•) LpL 0 μg/mL; (▪) 3 μg/mL; (▾) 12 μg/mL; *P < 0.05 vs. no added LpL at the same VLDL concentration. Kidney International 2001 59, 2062-2068DOI: (10.1046/j.1523-1755.2001.00720.x) Copyright © 2001 International Society of Nephrology Terms and Conditions

Figure 3 Effect of heparanase digestion on MC proliferation. Cells were quiesced as in Figure 2. Cells were stimulated for 24 hours with either 10% fetal calf serum or 5 μg/mL VLDL ± 5 μg/mL LpL. Heparanase is defined as cells treated with 1.0 U/mL bacterial heparanase for one hour prior to stimulation. Symbols are: () + heparanase; (□) control; *P < 0.05 vs. rest. Kidney International 2001 59, 2062-2068DOI: (10.1046/j.1523-1755.2001.00720.x) Copyright © 2001 International Society of Nephrology Terms and Conditions

Figure 4 MC platelet-derived growth factor (PDGF) gene expression in response to VLDL and LpL. Cells were quiesced as in Figure 2 and stimulated as in Figure 3 for eight hours. Quantitative PCR was performed using specific primers for PDGF-A chain and the constitutive control G3PDH. Results are expressed as ratio of PDGF to G3PDH. N = 4 to 6 per group. Symbols are: (□) control; () + heparanase; *P < 0.05 vs. rest. Kidney International 2001 59, 2062-2068DOI: (10.1046/j.1523-1755.2001.00720.x) Copyright © 2001 International Society of Nephrology Terms and Conditions