FIGURE 3.

The induction of Prx-2 is essential for the neuroprotective effect of Klotho. HT22 cells were infected with the control vector and four different constructs of shRNA. The Prx-2 expression was assessed by WB (A). HT22 cells stably infected with control vector or Prx-2 shRNA3 were pretreated with Klotho (KL) for 4 h and challenged with glutamate at the indicated concentrations. The extent of cytotoxicity was assessed 24 h later using CellTiter Glo (B). Asterisks indicate statistical significance of Klotho-treated versus Klotho-untreated samples: *, p < 0.05; **, p < 0.01 by Student's t test. Error bars, S.D. Results are representative of three independent experiments. C, qRT-PCR analysis of RNA obtained from whole brain samples of WT, KL-KO, and KL-OE mice. RNA was collected, and the mRNA analysis was performed using selective gene sets and normalized to endogenous controls. Each experiment was repeated at least two times, and representative results are shown.