Volume 70, Issue 9, Pages 1599-1606 (November 2006) The role of purinergic P2X7 receptors in the inflammation and fibrosis of unilateral ureteral obstruction in mice  R.G. Gonçalves, L. Gabrich, A. Rosário, C.M. Takiya, M. L.L. Ferreira, L.B. Chiarini, P.M. Persechini, R. Coutinho-Silva, M. Leite  Kidney International  Volume 70, Issue 9, Pages 1599-1606 (November 2006) DOI: 10.1038/sj.ki.5001804 Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 1 Histologic findings of kidney sections after 14 days of UUO stained by (a and b) hematoxylin–eosin and (c and d) periodic-acid Schiff, and (e and f) immunofluorescence for TUNEL. (a), (c), and (e) are from UUO WT mice and (b), (d), and (f) are from UUO P2X7 (−/−) mice. (a) Arrows point to increased cellularity and edema in the interstitium. (c) and (d) Arrows point to atrophic tubules. (e) and (f) Nuclei are all stained blue with apoptotic nuclei counterstained fluorescent green and identified with white arrowheads. Original magnification × 40. Kidney International 2006 70, 1599-1606DOI: (10.1038/sj.ki.5001804) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 2 (a and b) Immunohistochemical findings from kidney sections after 14 days of UUO by anti-f4/80, (e and f) anti-α-SMA (g and h), and anti-TGF-β antibodies. (c and d) show histological findings with Sirius-red staining. (a), (c), (e), and (g) are UUO WT mice; (b), (d), (f), and (h) are UUO P2X7 (−/−) mice. Original magnification × 40. Kidney International 2006 70, 1599-1606DOI: (10.1038/sj.ki.5001804) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 3 (a) The f4/80-positive cells in renal interstitium, (b) the α-SMA surface density and, (c) the TGF-β surface density in mice kidneys after 7 days (□) and 14 days (▪) of UUO. (a) *P<0.001 vs other groups; #P<0.001 vs UUO P2X7 (−/−) and ##P<0.001 vs other groups. (b) *P<0.01 vs other groups; #P<0.01 vs UUO P2X7 (−/−) and ##P<0.001 vs other groups. (c) *P<0.001 vs UUO P2X7 (−/−), **P<0.001 vs other groups; #P<0.001 vs UUO P2X7 (−/−) and ##P<0.001 vs other groups, by analysis of variance. Values are mean±s.d. Kidney International 2006 70, 1599-1606DOI: (10.1038/sj.ki.5001804) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 4 (a) The Sirius-red stained area, (b) the number of atrophic tubules per field and number of apoptotic cells, identified by (c) TUNEL technique in mice kidneys after 7 days (□) and 14 days (▪) of UUO. (A) *P<0.001 vs UUO P2X7 (−/−), **P<0.001 vs other groups, #P<0.001 vs UUO P2X7 (−/−) and ##P<0.001 vs other groups. (b) *P<0.001 vs other groups; #P<0.001 vs UUO P2X7 (−/−) and ##P<0.001 vs other groups. (c) *P<0.001 vs UUO P2X7 (−/−), **P<0.001 vs other groups; #P<0.001 vs UUO P2X7 (−/−) and ##P<0.001 vs other groups by analysis of variance. Values are mean±s.d. Kidney International 2006 70, 1599-1606DOI: (10.1038/sj.ki.5001804) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 5 Representative pictures of immunohistochemical study for anti-P2X7 antibody. (a) P2X7-positive epithelial cells of the cortex of UUO WT mice at day 7 of obstruction. No positive cells were detected in the interstitium. (b) Kidney cortex of UUO WT mice at day 14, with no positivity for anti- P2X7. Kidney International 2006 70, 1599-1606DOI: (10.1038/sj.ki.5001804) Copyright © 2006 International Society of Nephrology Terms and Conditions