Fig. 5.

(A) Astrocyte-rich cultures were transiently transfected for 24 h with the ARE reporter gene vector along with a Renilla luciferase expression vector. Transiently transfected cells were treated for 24 h with MCM₁₀ in the presence or absence of the Akt inhibitor Ly294002 (10 µM). MCM₁₀ induced a reduced ARE-related transcription activity, expressed by a lower luciferase activity. This effect was enhanced when the Akt signalling pathway was inhibited. Statistics: **p<0.01 vs control; ***p<0.005 vs control. (B) Transiently transfected cells were treated for 24 h with MCM₁₀ in the presence or absence of the GSK3β inhibitor LiCl (5 mM). Inhibition of GSK3β signalling pathway was able to fully reverse the effects of MCM₁₀ on the ARE-related transcription activity. Statistics: *p<0.05 vs control; ###p<0.005 vs MCM₁₀. (C) Transiently transfected cells were treated for 24 h with MCM₁₀ in the presence or absence of the p38 MAPK inhibitor SB203580 (20 µM). Inhibition of p38 MAPK signalling pathway fully reversed the effects of MCM₁₀ on the ARE promoter activity. Statistics: *p<0.05 vs control; ###p<0.005 vs MCM₁₀. (D) Transiently transfected cells were treated for 24 h with MCM₁₀ in the presence or absence of the GSK3β inhibitor LiCl (5 mM), the p38 MAPK inhibitor SB203580 (20 µM) or the two inhibitors combined. Inhibition of both GSK3β and p38 MAPK signalling pathways had additive effects and was able to fully reverse the effects of MCM₁₀ on the ARE promoter activity. Statistics: *p<0.05 vs control; ###p<0.005 vs MCM₁₀; +++p<0.05 vs MCM₁₀ + SB203580 or MCM₁₀ + LiCl.