Figure 2.

SHH Expression Was Downregulated in Stem Cells with Aging Both In Vitro and In Vivo

Sonic hedgehog (SHH) is considered a morphogen that regulates embryonic development. (A) In addition to this canonical function, SHH may act as an endogenous anti-aging factor. (B) Stem cells from mouse adipose tissue were isolated from young (5-week-old) and aged (72-week-old) mice, and then both mRNA and protein levels of SHH were evaluated using real-time PCR and western blotting, respectively. (C) After albumin and immunoglobulin depletion, the proteins in the serum samples from young and aged mice were precipitated with 10% TCA and subjected to western blotting using an anti-SHH polyclonal antibody. To prove media samples are not contaminated with cytosolic or nuclear content during TCA precipitation procedure, the levels of actin in the culture medium were analyzed. Brain (D), kidney (E), liver (F), and uterus (G) were isolated from young (5-week-old) and aged (72-week-old) mice, and then both mRNA and protein levels of SHH were evaluated using real-time PCR and western blotting, respectively. (H) The GEO database (https://www.ncbi.nlm.nih.gov/geo/) was analyzed to further verify the decreased SHH expression with aging. β-actin was used as an internal control. The results represent the means ± SD from three independent experiments.