FIGURE 5.

Klotho induces the phosphorylation of Akt. Rat primary hippocampal neurons were pretreated with Klotho (KL) for 4 h followed by additional treatment for 1 h (A), 24 h (B), and 48 h (C), in the absence or presence of glutamate (Glu) (3 mm). The lysates were collected for WB analysis using primary antibody recognizing the Thr³⁰⁸ phosphorylated site (top panels) and Ser⁴⁷³ phosphorylated site (bottom panels). Phosphorylated Akt proteins are normalized to total Akt (tAkt) control and presented as -fold increase normalized to untreated control. Asterisks indicate statistical significance of Klotho-treated versus Klotho-untreated samples. *, p < 0.05; **, p < 0.01; #, p < 0.05 for cells treated with glutamate only versus untreated cells by Student's t test. Error bars, S.D. The average of three independent experiments and representative blots are shown.