Figure 5. Robo2 deficiency protects mice from DOCA-salt-uninephrectomy–induced podocyte loss.

(A) Representative images of periodic acid-Schiff (PAS) staining of mouse kidney tissues show proteinaceous casts (asterisks), glomerulosclerosis, and increased matrix expression after 5-week deoxycorticosterone acetate (DOCA)-salt-uninephrectomy hypertension injury in wild-type, Myh9 conditional knockout (cKO), Robo2 cKO, and Myh-Robo2 double KO mice. Magnification: ×400. g, glomerulus. (B) Quantification of percentage of sclerotic glomeruli in kidney samples from A. Data are presented as the mean ± SEM, n = 3 animals per group, minimum 100 glomeruli counted in each animal. P = 0.028 compared with the Myh9 cKO group by 1-way ANOVA. (C) Representative images of WT1-positive podocytes in the glomeruli isolated from 4 groups of mice with Myh9/Robo2 single cKO, Myh9-Robo2 double cKO (dKO), and wild-type (WT) genotype after 5-week DOCA-salt-uninephrectomy injury. Magnification: ×400. g, glomerulus. (D) Quantification of podocyte number per glomerulus. Data are presented as the mean ± SEM, n = 3 animals per group, minimum 50 glomeruli counted in each animal. *P < 0.05, **P < 0.01 by 1-way ANOVA. (E) Albuminuria from the mice 5 weeks after DOCA-salt-uninephrectomy injury detected by acrylamide gel and Coomassie blue staining. WT, wild-type; Robo2 cKO, Robo2^fl/fl;Nphs2-Cre⁺ single knockout mice; Myh9 cKO, Myh9 ^fl/fl;Nphs2-Cre⁺ single knockout mice; dKO, Robo2^fl/fl;Myh9 ^fl/fl;Nphs2-Cre⁺ double knockout mice. (F) Quantification of urine albumin/creatinine ratios from each group of mice. Data are presented as the mean ± SEM, n = 4 animals. *P < 0.01, **P < 0.05 by 1-way ANOVA.