Figure 6.

SIRT1 activators affect HAS2–AS1 expression and NF-κB nuclear translocation. A, quantitative RT-PCR analyses are shown for HAS2–AS1 expression in AoSMCs treated for 24 h with 0.1 μg/ml TNFα alone or with 1 μm SRT1720 or 100 μm RESV. Data are expressed as relative expression of HAS2–AS1 with respect to its control (CNTR). Values are reported as mean ± S.E. of three independent experiments performed in duplicates. *, p < 0.05; **, p < 0.01; ***, p < 0.001. B, quantitative RT-PCR analyses are shown for AoSMCs nucleofected with 50 nm scrambled siRNA or HAS2 siRNA and left untreated or treated with 0.1 μg/ml TNFα for 24 h. Data are shown as mean ± S.E. of three independent experiments. *, p < 0.05; **, p < 0.01. C, representative images are shown for AoSMCs nucleofected with 4 μg of pcDNA3-GFP-RelA plasmid and grown on coverslips. Twenty four hours after the transfection, cells were treated as indicated for 24 h, washed in PBS, and observed by confocal microscopy (×63 objective). Bars, 20 μm.