Figure 9.

Determination of PI3K/AKT/GSK3β and Tau phosphorylation in vitro. A Tau phosphorylation cell model was established by treating SH-SY5Y cells with 10 μM Aβ1-42, and the cells were treated without or with 4c (0.1 μg/ml) or Evo (0.1 μg/ml) for 24 h. (A) The phosphorylation of p-PI3K, p-AKT, p-GSK-3β, and Tau in SH-SY5Y cells was detected by western blot using antibodies recognizing AT-8, T181, and Tau5 (n = 3). (B–F) The phosphorylation ratios compared with total protein were estimated by ImageJ (n = 3). (G, H) Tau phosphorylation was detected by immunofluorescent staining with anti-T181 antibody (n = 9 fields). Data are represented as mean ± SEM; ^*P < 0.05, ^**P < 0.01, and ^***P < 0.001.