Figure 4. Tubule-specific JAML deletion in mice only slightly ameliorates renal IRI.

(A and B) Experimental scheme for generating conditional knockout mice in which Jaml is specifically ablated in renal tubular cells (Ksp-Cre⁺ Jaml^fl/fl) by using Cre-loxP recombination system. Exon 4 is deleted upon Ksp-Cre–mediated recombination (A). Genotyping was confirmed by tail preparation and PCR at 2 weeks of age (n = 8) (B). (C) Representative Western blot and quantification of JAML expression in isolated tubules from Ksp-Cre⁺ Jaml^+/+ and Ksp-Cre⁺ Jaml^fl/fl mice (n = 4). (D) Representative IHC images of JAML in kidneys from Ksp-Cre⁺ Jaml^+/+ and Ksp-Cre⁺ Jaml^fl/fl mice with IRI (n = 5). Red arrows indicate representative JAML positive interstitial cells; black arrows indicate renal parenchymal cells. (E) SCr concentration in different groups of mice (n = 14). (F) BUN levels of different groups of mice (n = 14). (G) Representative images of H&E staining and quantitative assessment of tubular damage in the kidney from different groups of mice (n = 8). (H) Representative images and quantifications of IHC staining of KIM-1 in the kidney from different groups of mice (n = 8). Scale bar: 20 μm. Data are mean ± SEM. **P < 0.01, ***P < 0.001. Two-way ANOVA test (E–H), 2-tailed Student’s unpaired t test analysis (C). See complete unedited blots in the supplemental material.