FIG. 3.

Peroxide-induced autophagy is associated with inactivation of the mTOR pathway and 3MA inhibits autophagy but not transient phosphorylation of Akt. (A) Western blotting of LC3 in SH-SY5Y cells exposed to 200μM H₂O₂ (perox) for up to 4 h. As a positive control, the homogenate of SH-SY5Y cells exposed to 100nM rapamycin (rap) for 30 min is included. The filter was tripped and reprobed for actin. The LC3 II/actin ratio was calculated by densitometry of three independent experiments. Statistical significance between relevant treatments is indicated. (B) Western blotting of ser235/236-pS6 in SH-SY5Y cells treated as for panel (A). The filter was tripped and subsequently reprobed for total S6 and for tubulin. The pS6/S6 ratio was calculated by densitometry of three independent experiments. Statistical significance between relevant treatments is indicated. (C) Western blotting of LC3 in SH-SY5Y cells exposed to 200μM H₂O₂ (perox) for 2 h in the absence or the presence of 3MA. The LC3 II/actin ratio was calculated by densitometry of three independent experiments. Statistical significance between relevant treatments is indicated. (D) Western blotting of ser273-pAkt in SH-SY5Y cells exposed to 200μM H₂O₂ (perox) for 30 min in the absence or the presence of 3MA. The pAkt/Akt ratio was calculated by densitometry of three independent experiments. Statistical significance between relevant treatments is reported (n.s., not significant).