Fig. 5.

High Klotho is less effective in attenuating ischemia-reperfusion injury in mice with low beclin 1 activity. Transgenic mice overexpressing Klotho (Tg-Kl mice) and their wild-type (WT) littermates, Becn1^+/− and chimeric mice with Tg-Kl (Becn1^+/−;Tg-Kl), and Bcl2^AAA mice and chimeric mice with Tg-Kl (Bcl2^AAA;Tg-Kl) were subjected to IRI operations. Two days after surgery, mice were euthanized, blood was drawn, and kidneys were harvested. A: kidney function and plasma creatinine (Cr). B: kidney histology with hematoxylin and eosin staining. Scale bar = 100 μm. C: kidney histology with periodic acid-Schiff (PAS) staining. Top, representative microscopic images of PAS staining. Scale bar = 100 μm. Bottom, semiquantitative assessment of kidney injury score based on PAS stain in each group. D: kidney damage markers. Top, representative immunoblots for active caspase-3 and neutrophil gelatinase-associated lipocalin (NGAL) proteins in total kidney lysates. Bottom, summary of all immunoblots from each group. E: Klotho expression and autophagic markers in the kidney. Left, representative immunoblots for Klotho, beclin 1, Bcl-2, light chain (LC)3, and p62 in total kidney lysates. Right, summary of all immunoblots from each group. Data are expressed as means ± SD, and statistical significance was evaluated by two-way ANOVA followed by a Student-Newman-Keuls post hoc test and significance was accepted when *P < 0.05 and **P < 0.01 between two groups. Sample numbers in each group are presented in brackets underneath the corresponding bars.