Figure 3.

SIRT6 is required for repair via the BER pathway. (A) Transient overexpression of SIRT3 and SIRT6 in HCA2-hTERT cells. A vector for expressing SIRT3 or SIRT6 was transfected into HCA2-hTERT cells. At the indicated time points, cells were harvested for Western blot analysis of sirtuin expression. (B) Overexpression of SIRT6 promotes BER efficiency. A pcDNA3.1 control vector or a vector encoding SIRT3 or SIRT6 was co-transfected with damaged pEGFP-N1 and pDsRed2-N1. Seventy-2 hours later, cells were harvested for analysis of BER efficiency using FACSverse. (C) Western blot analysis of mSIRT6 in wild type and SIRT6 knockout MEFs. Exponentially growing MEFs were harvested for to quantify protein expression by Western blot analysis. (D) BER efficiency is reduced in SIRT6 knockout MEFs. 0.05 μg of MB+VL treated pEGFP-N1 and 0.005 μg pDsRed2-N1 were co-transfected into wild type or SIRT6 knockout MEFs 72 hours before cells were harvested for FACS analysis. All the experiments were repeated at least 3 times and error bars represent SD. ** P < 0.01