Fig. 3

Deletion of Aim2 gene attenuates the inflammatory response in aortic VSMC of mice. a-c Analysis of mRNA expression of Aim2 (a), Nlrp3 (b), Il1b (c), by real-time RT-PCR (n = 3 measurements of 3 experiments for each). Total RNA was extracted from VSMC grown in vitro with normal growth medium (−OM) or osteoblast mineralization medium (+ OM) at day 14. A.u. arbitrary unit; relative expression of target gene was normalized against expression of housekeeping genes (Actb, Gapdh and B2m). d, e Concentrations of secreted IL-1β (d) and IL-6 (e) in the supernatant of WT and Aim2−/− VSMC was determined by ELISA. Data show the mean and SEM of n = 3 independent VSMC pools analyzed in duplicate. a-e Data were tested by one-way ANOVA. ns: not significant, *: P < 0.05, **: P < 0.01, ****: P < 0.0001 f Protein expression of phosphorylated NF-kB and phosphorylated H2A.X were detected by Western blotting. Shown are representative examples of two VSMC pools derived from 2 to 4 animals each. Upper panel: numbers represent the ratio of phospho-NF-kB to total NF-kB, normalized to GAPDH expression in the same sample. Lower panel: numbers represent the ratio of phospho-H2A.X and GAPDH. The expression in proliferating WT VSMC was set as 1.0