Fig. 2

SIRT6 silencing accelerates Snail protein turnover. a Western blot analysis of Snail protein in A549 and H1299 cells transfected with control and SIRT6 shRNA. One representative experiment of three independent experiments are shown. b SIRT6-overexpressing A549 and H1299 cells were immunoprecipitated with anti-SIRT6 or control antibody and subjected to Western blot analysis of Snail, Twist1, and SIRT6. One representative experiment of three independent experiments are shown. c Analysis of Snail acetylation status. A549 and H1299 cells expressing SIRT6 were immunoprecipitated with anti-Snail antibody and probed with anti-acetyl lysine antibody. One representative experiment of three independent experiments are shown. d Effect of SIRT6 silencing on Snail acetylation status. A549 and H1299 cells transfected with control and SIRT6 shRNA were immunoprecipitated with anti-Snail antibody and probed with anti-acetyl lysine antibody. One representative experiment of three independent experiments are shown. e Cells were treated with cycloheximide (CHX) and tested for Snail protein levels at indicated time points. Data shown are mean from three independent experiments performed in triplicate. The Snail protein level at 0 h was assigned a value of 1. f Western blot analysis demonstrated that MG132 pretreatment blocked the downregulation of Snail protein in SIRT6-depleted A549 and H1299 cells. Data shown are mean from three independent experiments performed in triplicate. The Snail protein level at shCtrl-transfected cells was assigned a value of 1. ^*P < 0.05