Figure 6

Conditional depletion of macrophages in LysM-Cre/DTR mice using diphtheria toxin (DT) attenuates the MMT response and renal fibrosis in obstructive nephropathy (UUO). (a and b) Results of three-color confocal immunofluorescence show that administration of DT reduces F4/80+ macrophages by 60% in the UUO kidney, leading to a dramatic reduction in the accumulation of F4/80⁺α-SMA⁺ cells and total α-SMA⁺ myofibroblasts. (c) Immunohistochemistry off collagen I staining. (d) Real-time PCR analysis of α-SMA and collagen I mRNA levels; (e) western blot analysis and quantification of α-SMA and collagen I protein levels. Each bar represents mean±S.E.M. for six mice. *P<0.05, **P<0.01, ***P<0.001 versus sham-control; ^##P<0.01, ^###P<0.001 saline-control or as indicated. Scale bar, 50 μM