Figure 5. Inactivation of GSK3β enhanced pyknotic necrosis.

(A) Serum-starved SAS cells treated with 0.3 mg/ml ANE and with or without 5 mM NAC were harvested at the indicated time points for detecting GSK3β, p-GSK3β and cleaved PARP by Western blot. (B) Serum-starved cells treated with SB216763 in the presence of H₂O₂ or ANE were stained with AO/EtBr. (C) Similar to the condition in (B), cells were harvested for Western blot analysis 4 hours after treatment. (D) Viability of serum-starved cells treated with SB216763 in the presence or absence of 20 µM Z-VAD-fmk was measured using MTT assay 16 hours after treatment.