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. 2016 Apr 1;310(10):L964–L974. doi: 10.1152/ajplung.00054.2016

Fig. 2.

Fig. 2.

Sirt-2 inhibition reverses the repression of LPS-mediated NF-κB activation and target gene expression by 17-AAG. A: adeno-GFP and adeno-NF-κB reporter luciferase viral particles were cotransduced as in Fig. 1. Cells were treated with LPS for 4 h in the absence and presence of 5 μg/ml 17-AAG and 50 μM Sirtinol (Sirt-2 selective inhibitor, together for 16 h, pretreatment). Luciferase activity (± SE, n = 3) was significantly increased by LPS; 17-AAG suppressed the LPS effect, whereas Sirtinol potentiated the LPS effect and partially recovered the 17-AAG effect on LPS treatment in HLMVEC. B: TaqMan qRT-PCR analysis (± SE, n = 3) showed that LPS induced IKBα mRNA expression; 17-AAG suppressed the LPS effect, the sirtuin inhibitor potentiated the LPS and reversed the 17-AAG effect in HLMVEC. B (i), (ii), and (iii): 50 μM Sirtinol-, 10 μM AGK2-, and 10 μM EX527-treated cells, respectively. C: Sirt-2 expression was downregulated by Sirt-2 shRNA lentiviral particles in HLMVEC; cells were then treated with LPS in the presence and absence of 17-AAG. Bar graph (± SE) represents fold change in IKBα mRNA expression in 1 of 3 independent experiments. LPS induced IKBα mRNA expression in wild-type and potentiated IKBα mRNA expression in Sirt-2 knock-down HLMVEC; 17-AAG suppressed and partially but significantly reversed the effect of 17-AAG in Sirt-2 knock-down HLMVEC.