Fig. 4

KLF4 is upregulated in SIRT6-depleted NSCLC cells. a Real-time PCR analysis of indicated transcripts in A549 and H1299 cells transfected with control and SIRT6 shRNA. Mean fold change was calculated from three independent experiments performed in triplicate, with respect to shCtrl-transfected cells arbitrarily assigned a value of 1. ^*P < 0.05 vs. control cells. N.S. means no significance. b ChIP experiments using anti-Snail antibody showed that SIRT6 depletion suppressed the enrichment of Snail at the promoter of Klf4. Data shown are mean ± SD of three independent experiments performed in triplicate. The control group was arbitrarily assigned a value of 1. c Real-time PCR analysis of KLF4 mRNA levels in cells transfected with indicated constructs. Data shown are mean ± SD of three independent experiments performed in triplicate. Empty vector group was arbitrarily assigned a value of 1. d Western blot analysis of KLF4 protein levels in cells treated as in (c). One representative experiment of three independent experiments are shown. e ChIP experiments using anti-Snail antibody showed that SIRT6 overexpression promoted the enrichment of Snail at the promoter of Klf4. Data shown are mean ± SD of three independent experiments performed in triplicate. The control group was arbitrarily assigned a value of 1. ^*P < 0.05