Subcutaneous human BMMSC transplantation up-regulated Klotho expression in immunocompromised mice. (A) Immunohistochemical staining with anti-Klotho antibody showed that transplant recipient mice (Transplant) expressed higher level of Klotho () in the epithelial cells of the renal tubules (RT) than those of age-matched controls (Control). G indicates glomerulus. Original magnification ×200. Bars represent SD. (B) Western blot analysis confirmed that Klotho was elevated in kidney of transplant recipient (Transplant, n = 3) compared with nontreated littermates (Control, n = 3). (C) Likewise, Western blot analysis showed elevated Klotho in brain of transplant recipient mice (Transplant n = 3) compared with the Control (n = 3). (D) ELISA assay further confirms elevated serum Klotho level in transplant recipient mice (Transplant, n = 3) compared with the control group (Control, n = 3). Bars represent SD. (E) Random blood glucose measurement showed decreased serum glucose in recipient mice (Transplant, n = 3) in comparison to control mice (Control, n = 3). Bars represent SD. (F) Similarly, serum IGF-I level was lower in recipient mice (Transplant, n = 3) compared with control mice (Control, n = 3). Bars represent SD. (G) Urine protein level was significantly decreased in recipient mice (Transplant, n = 3) compared with control mice (Control, n = 3). Bars represent SD. (H) Western blot analysis showed that expression of insulin receptor (IR) α, IRβ, phosphatidylinositol 3-kinase (PI3-K) p85, and PI3-K p110 was down-regulated in kidney tissues of transplant recipient mice (Transplant, n = 3) compared with control mice (Control, n = 3). β-Actin was used as protein loading control.