(a) An experimental protocol for the study on the anti-GBM nephritis in tamoxifen-inducible systemic CTGF cKO (Rosa-CTGF cKO) mice. Three-week old male ROSA26-CreERT2; Ctgffl/fl mice or control [Cre (−); Ctgffl/fl] were intraperitoneally injected with 4-OHT, and anti-GBM nephritis was induced at 8 weeks of age (GBM). Vehicle treatment was also carried out in some animals (veh.). Mice were killed at 4 weeks after induction of anti-GBM nephritis. (b) Double immunofluorescence staining for CTGF (green) and podocin (red) in the glomeruli of control or Rosa-CTGF cKO mice at 4 weeks after induction of anti-GBM nephritis. Bar represents 50 μm. (c) Changes in proteinuria during the course of anti-GBM nephritis. Closed squares, vehicle-treated control mice; open square, vehicle-treated Rosa-CTGF cKO mice; closed circles, control mice with GBM nephritis; open circles, Rosa-CTGF cKO mice with anti-GBM nephritis. (d) Representative photomicrographs of the kidneys at 4 weeks after induction of anti-GBM nephritis (PAS staining). Left upper panel, vehicle-treated control mice; right upper panel, vehicle-treated Rosa-CTGF cKO mice; left lower panel, control mice with anti-GBM nephritis; right lower panel show, Rosa-CTGF cKO mice with anti-GBM nephritis. Bar represents 50 μm. (e) The percentage of crescent formation (left panel) and the mesangial area (right panel) in anti-GBM nephritis. (f) Electron microscopic analysis of glomeruli. Bar represents 2,000 nm. (g) Immunofluorescence studies for rabbit IgG, mouse IgG and mouse C3 deposition in the kidney at 28 days after induction of anti-GBM nephritis in control and in Rosa-CTGF cKO mice. Both Rosa-CTGF cKO and control mice exhibited liner IgG and C3 deposition along the GBM. Bar represents 50 μm. Values are expressed as means ± s.e. *P < 0.05, **P < 0.01 vs control mice with GBM nephritis. Vehicle-treated control mice (n = 7), vehicle-treated Rosa-CTGF cKO mice (n = 7), control mice with GBM nephritis (n = 7), Rosa-CTGF mice with GBM nephritis (n = 9). Veh, vehicle; GBM, anti-GBM nephritis.