Figure 7.

G93A-SOD1 toxicity is not mediated by p53 acetylation state or by IRS-2/Ras/ERK1/2 pathway in SH-SY5Y cells. (a) Western blot analysis of 20 μM of total protein extract from cells infected with adenoviral vectors coding for Wt-SOD1 (Wt) and G93A-SOD1 (G93A) and treated with 3 μM Ex527 or DMSO. Antibodies against SIRT1, Erk1/2, pErk1/2, p53, p53-Ac, tubulin and Ac-tubulin were used. β-Actin was used as loading control. One representative blot is shown from three independent experiments giving comparable results. (b) Densitometric analysis of results as in (a); data are expressed as acetylation ratio of p53 and tubulin. (c) Cells infected with adenoviral vectors coding for Wt-SOD1 (Wt) and G93A-SOD1 (G93A) were treated with 3 μM Ex527 or with 3 μM SL327 or both. Cell viability was assessed and reported for Figure 5a. Values significantly different from relative controls are indicated with *P<0.05 with respect to Ctrl and ^#P<0.05 with respect to DMSO. (d) Western blot analysis of 20 μM of total protein extract from C2C12 cells differentiated for 5 days and expressing G93A-SOD1 (G93A) or untransfected (Ctrl). Antibodies against SIRT1, p53 and p53-Ac were used. β-Actin was used as loading control. One representative blot is shown from three independent experiments giving comparable results. (e) Densitometric analysis of data as in (d); data are expressed as acetylation ratio of p53