Figure 6.

SIRT1 overexpression inhibits OPN-induced NF-κB activation.

Notes: A549 and NCI-H358 cells infected with LV-SIRT1 or LV-NC were treated with OPN (400 ng/mL) for 2 hours. (A) The levels of NF-κB p65 protein in cellular nucleus and cytoplasm were determined by Western blot analysis. Lamin B and β-Actin, respectively, indicate the nuclear and cytoplasmic fraction. The group treated with CB + LV-NC serving as control is presented as 1. Results presented are mean ± SD analyzed with Student’s t-test from three independent experiments. *P<0.05, ***P<0.001 versus control. ^#P<0.05, ^##P<0.01 versus OPN + LV-NC. (B) The locations of NF-κB p65 in the cells were examined by immunofluorescence. The group treated with CB + LV-NC served as control. The cells were immunostained with anti-NF-κB p65 antibody (green) and the nucleus was stained with DAPI. Scale bars represent 50 μm. Magnification ×100.

Abbreviations: SIRT1, Sirtuin 1; OPN, osteopontin; NF-κB, nuclear factor-κB; LV-SIRT1, lentiviral vectors overexpressing SIRT1; NC, negative control; CB, control buffer; SD, standard deviation.