Figure 5.

REST protects against doxorubicin-induced nephropathy. (A) Representative urine Coomassie blue staining 7 days after venous retro-orbital injection of either vehicle or a single dose of 17 μg/g of doxorubicin in BALB/c WT mice. BSA is shown as a positive control. (B) Trichrome staining of the kidneys 7 days after retro-orbital injection of either vehicle (n=6) or a single dose of 17 μg/g of doxorubicin into Balb/c WT mice. Scale bar, 10 μm. (C) Nephrin and WT1 mRNA in the kidneys of mice injected with either vehicle or a single dose of 17 μg/g of doxorubicin. (D) Western blot (left) and quantification (right) of REST cleaved PARP, cleaved caspase-3 (CC3) (top blot), and nephrin (lower) 7 days after injection of either vehicle or a single dose of 17 μg/g of doxorubicin. (E) Periodic acid–Schiff staining (left) and quantitative analysis of glomerular lesion scores (right) in kidneys of C57bl/6 REST^flox/flox and REST^Δpod mice, 3 months after injection of either vehicle or a single dose of doxorubicin (17 μg/g). At the time of sacrifice, the kidneys of vehicle-injected mice and REST^flox/flox mice injected with doxorubicin had no obvious abnormal histology, whereas REST^Δpod mice injected with doxorubicin exhibited glomerular lesions with glomerulosclerosis, tubular vacuolization and intratubular casts. Scale bar, 10 μm. (F) REST^Δpod mice injected with doxorubicin displayed albuminuria compared with doxorubicin treated REST^flox/flox mice. (G) Western blot (upper panel) and quantification of nephrin and synaptopodin (lower panels) in C57bl/6 REST^flox/flox and REST^Δpod mice, injected with either vehicle or a single dose of doxorubicin (17 μg/g). *P<0.05, **P<0.01, ***P<0.001, ***P<0.0001 between groups, ANOVA, and post-hoc Tukey–Kramer test were used.