Fig. 9.

a, b The effect of necrostatin-1 (Nec-1) on H₂O₂-induced cathepsin D activity in UN- (a) and RA (b) SH-SY5Y cells. The cells were pre-treated for 30 min with Nec-1 (1–40 μM) or pepstatin A (PsA; 0.2 μM) followed by 18 h of treatment with H₂O₂ (0.25 and 0.5 mM for UN- and RA-SH-SY5Y cells, respectively). Data from duplicate determinations in 3–4 independent experiments were normalized to the protein level and are expressed as percentages of the control ± SEM. c, d The effect of necrostatin-1 (Nec-1) on H₂O₂-induced cathepsin D expression in UN- (c) and RA- (d) SH-SY5Y cells. The cells were pre-treated for 30 min with Nec-1 (20 μM) followed by 18 h of treatment with H₂O₂ (0.25 and 0.5 mM for UN- and RA-SH-SY5Y cells, respectively). Expression of 43 and 33 kDa forms of cathepsin D was done by Western blot method. Histograms show the quantified WB results from duplicate determinations in 2 independent experiments which were normalized to the protein loading control (GAPDH) and are expressed as fold of the control ± SEM