FIG. 8.

Effect of APAP-CM from peritoneal macrophages from wild-type and SIRT1-Tg mice in the modulation of SIRT1 protein levels in hepatocytes. (A) Peritoneal macrophages were isolated from SIRT1-WT and SIRT1-Tg mice and then used to prepare control-CM or APAP-CM as previously described. Then, mouse hepatocytes were treated with these CM for 16 h. Immunoblots showing SIRT1 protein levels and α-Tubulin as a loading control. Densitometric quantification showing SIRT1 protein levels in each condition. Values are mean ± SEM. Statistical analysis was performed by two-way ANOVA followed by Bonferroni post hoc test. *p < 0.05 versus control-CM. (B) Levels of the active fragment of caspase-1 (p10) detected in peritoneal macrophages derived from SIRT1-WT and SIRT1-Tg mice stimulated with APAP (5 mM) for 3 or 6 h. GAPDH showing similar amounts of protein loaded in each lane. Representative blots are shown. After quantification of all blots, results are expressed as fold change relative to vehicle-treated SIRT1-WT macrophages and are mean ± SEM. Statistical analysis was performed by two-way ANOVA followed by Bonferroni post hoc test. *p < 0.05 versus wild-type macrophages. GAPDH, glyceraldehyde-3-phosphate dehydrogenase.