Figure 4.

Protein expression patterns of HDAC5, HDAC11, SIRT1 and SIRT2 in differentiated human SH-SY5Y neuroblastoma cells. SH-SY5Y cells were uninfected (Ctrl) or infected with adenoviral vectors coding for wild-type SOD1 (Wt) or G93A-SOD1 (G93A). (a) Western blot analysis of 20 μg of cell lysate using antibodies against HDAC5, HDAC11, SIRT1, SIRT2, p53-Ac and Ac-tubulin. β-Actin was used as loading control, SOD1 as infection control, and P-53 and tubulin to monitor the acetylation rate. (b) Densitometric analysis of n=3 experiments as in (a). Values significantly different from relative controls are indicated with **^,##P<0.01. (c and d) Western blot analysis to detect PGC1α and p53 acetylation, respectively, in the immunoprecipitate with anti Ac-lysine antibody. In the lower panel, 5% of input is shown. (e) Immunolocalization of HDAC5, HDAC11, SIRT1 and SIRT2. Panels show typical images observed in n=3 independent experiments. Scale bar: 5 μm