Figure 4

Involvement of GSK-3β/HSF-1-dependent HSP expression. (a) Cells were pretreated with Ntn-1 (10 ng/ml) and/or Akt inhibitor (10 μM) for 30 min before being exposed to hypoxia for 72 h; phospho-GSK-3β and GSK-3β were detected by western blot. (b) Cells were pretreated with Ntn-1 or LiCl (10 mM) for 30 min before being exposed to hypoxia for 72 h; phospho-GSK-3β and GSK-3β were detected by western blot. (c) Cells were pretreated with Ntn-1 or LiCl (10 mM) for 30 min before being exposed to hypoxia, and HSF-1, lamin A/C, pan-cadherin, and β-actin in the nuclear and non-nuclear fraction were detected by western blot. Each of the examples (a–c) is representative of four independent experiments. The right part (a–c) depicting the bars denotes the mean±S.E. of four independent experiments for each condition determined from densitometry relative to β-actin or total GSK-3β. *P<0.05 versus control, **P<0.05 versus hypoxia alone, ^#P<0.05 versus combination of hypoxia and Ntn-1. ROD, relative optical density