Effects of the time course of i.t. Ad-PTEN on the spinal PTEN and phospho-mTOR levels in CCI rats. (A) Western blots for PTEN, phospho-mTOR, and β-actin proteins from sham-operated plus i.t. vehicle 7 days post injury, CCI plus i.t. Ad-GFP 7 days post injury, CCI plus i.t. Ad-PTEN 7 days post injury, sham-operated plus i.t. vehicle 14 days post injury, CCI plus i.t. Ad-GFP 14 days post injury, and CCI plus i.t. Ad-PTEN 14 days post injury. (B) Relative density of the immunoblot of PTEN. (C) Relative density of the immunoblot of phospho-mTOR. Relative band intensities of PTEN and phospho-mTOR were quantified by densitometry and indicated as the percent change relative to that for the sham-operated 7 days post injury group (100%). Western blotting revealed that PTEN and phospho-mTOR protein expression were detectable in the sham-operated group, but CCI-induced downregulation of PTEN and upregulation of phospho-mTOR protein on day 7 and day 14 after CCI surgery were both attenuated by i.t. Ad-PTEN. Western blotting of β-actin was performed to verify that equivalent amounts of protein were loaded in each lane. Each bar in (B) and (C) represents the mean ± SEM with three rats per group. *P < 0.05 compared with sham-operated 7 days post injury group. #
P < 0.05 between CCI plus i.t. Ad-PTEN group compared with the same time points in CCI plus i.t. Ad-GFP group. Ad-GFP, adenovirus-mediated green fluorescent protein; Ad-PTEN, adenovirus-mediated phosphatase and tensin homolog deleted from chromosome 10; CCI, chronic constriction injury; p-mTOR, phosphorylated mammalian target of rapamycin.