Over expression of SIRT6 or H133W.

<p>HEK293 cells were transfected with plasmid expressing Flag-tagged wild type SIRT6 (lanes 3 and 7) or deacetylase-dead (H133W) mutant of human SIRT6 in (lanes 4 and 8). Control cells were transfected with no DNA (lanes 1 and 5) or empty vector DNA pCDNA3 only (lanes 2 and 6). Each lane was loaded with 50 µg total protein of cell extract and analysed by SDS-PAGE followed by Western blotting with either anti-SIRT6 or anti-Flag. RNA was extracted from these cells was used for transcriptomic analysis without and with TNFα stimulation for 4 h.