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. 2018 Dec 4;33(3):3704–3717. doi: 10.1096/fj.201800321R

Figure 1.

Figure 1

A–C) SIRT6 expression modulates the intracellular NAD(H) levels and NAMPT activity. BxPC-3 (A) and MCF-7 (B, C) cells were engineered by retroviral transduction to express the WT form of SIRT6 (SIRT6 WT), or the catalytically inactive SIRT6 mutant (SIRT6H133Y), or were transduced with the empty vector pBP; the same cells were engineered to down-regulate SIRT6 expression (sh2-SIRT6) or were transduced with the respective empty vector pRS. Cells were lysed, and intracellular NAD+ and NADH levels were determined (A, B). The mean ± sd of ≥4 independent determinations. Cells were lysed, and SIRT6 expression level was evaluated by Western blot analysis (representative results are shown) (C). D, E) BxPC-3 cells expressing different levels of SIRT6 were lysed and NAD+ synthesis was evaluated by adding NAM and PRPP as substrates (D), and the iNAMPT expression level was evaluated by Western blot analysis (E). The mean ± sd of results of 3 different analyses, and 1 representative Western blot are shown. *P < 0.05, **P < 0.01, ***P < 0.001 vs. respective controls.