Figure 2.DNA damage stabilizes SIRT6 interaction with chromatin.(a)
Gel shift showing SIRT6, but not SIRT1, binding to purified native
nucleosomes in vitro. Left: coomassie stain of recombinant SIRT6 and SIRT1
proteins. Right: EtBr stain of nucleosomal DNA on a non-denaturing gel;
arrow indicates SIRT6-mononucleosome complex. (b) Schematic of
chromatin purification protocol (see methods). Dt, detergent extractable
fraction; Rn, RNase extractable fraction; Chr, purified chromatin
fraction. (c) DNA damage-dependent stimulation of SIRT6 association
with chromatin. Western analysis with the indicated antibodies of HeLa
cells treated with 45 nM neocarzinostatin (NCS) for 1 hour and fractionated
as described in (c). H4 and Tubulin are detected in the expected
fractions.
