Figure 1.
IL-6 is induced in the kidney following obstructive injury.
A. Graphic presentation shows IL-6 mRNA induction. ** P<0.01 vs normal control kidney. n = 4. B. Representative photomicrographs of kidney sections stained for IL-6 (brown) and counter stained with hematoxylin (blue) (Original magnification: X400).
Figure 2.
Effect of IL-6 deficiency on the accumulation of bone marrow-derived fibroblast precursors in the kidney after UUO.
A. Representative photomicrographs of kidney sections from WT and IL-6 KO mice 5 days after UUO stained for CD11b (red), procollagen I (green), and DAPI (blue). B. Quantitative analysis of CD11b+ and procollagen I+ fibroblasts in the kidney of WT and IL-6 KO mice 5 days after UUO. ** P<0.01 vs WT control, # P>0.05 vs WT-UUO, and ++ P<0.01 vs KO-UUO. n = 4 per group. C. Representative photomicrographs of kidney sections from WT and IL-6 KO mice 1 week after UUO stained for CD11b (red), procollagen I (green), and DAPI (blue). D. Quantitative analysis of CD11b+ and procollagen I+ fibroblasts in the kidney of WT and IL-6 KO mice 1 week after UUO. ** P<0.01 vs WT control, # P>0.05 vs WT-UUO, and ++ P<0.01 vs KO-UUO. n = 5 per group. E. Representative cytometric diagrams showing the effect of IL-6 deficiency on the accumulation of CD11b and collagen I dual positive fibroblasts in the kidney of WT and IL-6 KO mice 1 week after UUO. F. Quantitative analysis of CD11b and collagen I dual positive fibroblasts in the kidney of WT and IL-6 KO mice 1 week after UUO. ** P<0.01 vs WT control, # P>0.05 vs WT UUO, and ++ P<0.01 vs KO UUO. n = 3 per group.
Figure 3.
Effect of IL-6 deficiency on myofibroblast activation and α-SMA expression in obstructive nephropathy.
A. Representative photomicrographs of α-SMA immunostaining in the kidney of WT and IL-6 KO mice after UUO. B. Quantitative analysis of α-SMA protein expression in the kidney of WT and IL-6 KO mice after UUO. ** P<0.01 vs WT controls; # P>0.05 vs WT UUO; ++ P<0.01 vs KO UUO. n = 5 per group. C. Representative Western blots show the levels of α-SMA protein expression in the kidney of WT and IL-6 KO mice. D. Quantitative analysis of α-SMA protein expression in the kidney of WT and IL-6 KO mice. ** P<0.01 vs WT controls; # P>0.05 vs WT UUO; + P<0.05 vs KO UUO. n = 3 per group.
Figure 4.
Effect of IL-6 deficiency on profibrotic molecule expression.
A. IL-6 deficiency does not affect CXCL16 gene expression. The mRNA levels of CXCL16 in the kidneys of WT and IL-6 KO mice were determined by real-time RT-PCR. ** P<0.01 vs WT control, # P>0.05 vs WT UUO, and + P<0.05 vs KO UUO. n = 3–4 per group. B. IL-6 deficiency does not influence TGF-β1 gene expression. The mRNA levels of TGF-β1 in the kidneys of WT and IL-6 KO mice were determined by real-time RT-PCR. ** P<0.01 vs WT control, # P>0.05 vs WT UUO, and + P<0.05 vs KO UUO. n = 3–4 per group. C. IL-6 deficiency does not influence IL-4 gene expression. The mRNA levels of IL-4 in the kidneys of WT and IL-6 KO mice were determined by real-time RT-PCR. ** P<0.01 vs WT control, # P>0.05 vs WT UUO, and + P<0.05 vs KO UUO. n = 3–4 per group. D. IL-6 deficiency does not influence IL-13 gene expression. The mRNA levels of IL-13 in the kidneys of WT and IL-6 KO mice were determined by real-time RT-PCR. ** P<0.01 vs WT control, # P>0.05 vs WT UUO, and + P<0.05 vs KO UUO. n = 3–4 per group.
Figure 5.
Effect of IL-6 deficiency on renal fibrosis and extracellular matrix deposition in the kidney.
A. Representative photomicrographs show kidney sections stained with picrosirius red for assessment of total collagen deposition. B. Bar graph shows quantitative analysis of renal interstitial collagen in different groups as indicated. ** P<0.01 vs WT control, & P>0.05 vs WT UUO, and ++ P<0.01 vs KO UUO. n = 5 per group.
Figure 6.
Effect of IL-6 deficiency on collagen I expression in the kidney.
A. Representative photomicrographs of collagen I immunostaining in the kidney of WT and IL-6 KO mice after surgery (original magnification X400). B. Quantitative analysis of interstitial collagen I protein expression in the kidney sections of WT and IL-6 KO mice. ** P<0.01 vs WT-control, # P>0.05 vs WT UUO, and ++ P<0.01 vs KO UUO. n = 5 per group. C. Representative Western blots show the protein levels of collagen I in the kidney of WT and IL-6 KO mice. D. Quantitative analysis of collagen I protein expression in the kidney of WT and IL-6 KO mice. ** P<0.01 vs WT controls, # P>0.05 vs WT UUO, and + P<0.05 vs KO UUO. n = 4 per group.
Figure 7.
Effect of IL-6 deficiency on fibronectin expression in the kidney.
A. Representative photomicrographs of fibronectin immunostaining in the kidney of WT and IL-6 KO mice after UUO (original magnification X400). B. Quantitative analysis of interstitial fibronectin protein expression in the kidney sections of WT and IL-6 KO mice. ** P<0.01 vs WT control, # P>0.05 vs WT UUO, and ++ P<0.01 vs KO UUO. n = 5 per group. C. Representative Western blots show the protein levels of fibronectin in the kidney of WT and IL-6 KO mice. D. Quantitative analysis of fibronectin protein expression in the kidney of WT and IL-6 KO mice. ** P<0.01 vs WT controls, # P>0.05 vs WT UUO, and + P<0.05 vs KO UUO. n = 4 per group.