Figure 2. Generation and characterization of P3h2^ΔPod mice by urinalysis.

(A) Schematic strategy for the generation of P3h2^ΔPod mice. (B) KO confirmation of the P3h2^ΔPod mice. qPCR P3h2 mRNA expression analysis was performed in sorted podocytes from WT and KO mice. P3h2 mRNA levels were reduced by 99% in P3h2^ΔPod mice compare with levels in P3h2^fl/fl mice. (C) Immunofluorescence staining of WT and KO kidney tissues for P3H2, NPHS1, and DAPI. There was no detectable P3H2 in the P3h2^ΔPod mouse podocytes. Scale bar: 10 μm; inset zoom, ×5. (D) Body weights of P3h2^ΔPod and P3h2^fl/fl mice were measured starting at 5 weeks until 48 weeks of age. There was no significant difference at any time point in body weights between the WT and KO mice (E) UACR of P3h2^ΔPod and P3h2^fl/fl mice. KO mice started to present with albuminuria at 36 weeks of age, and this had increased at 48 weeks of age. (F) Serum urea measurements for P3h2^ΔPod and P3h2^fl/fl mice. No significant increase was observed in the KO mice. (G) Serum cystatin C measurement for P3h2^ΔPod and P3h2^fl/fl mice. No significant increase was observed in the KO mice. (H) Hematuria was detected in spot urine of P3h2^ΔPod mice. Representative images of urine from mice of each genotype. There were significantly more and dysmorphic RBCs in KO urine than WT urine. Scale bar: 50 μm: inset zoom, ×5. (I) Quantification of urinary RBCs under a light microscope. n ≥ 3. Graphs show the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001, by unpaired, 2-tailed Student’s t test.