Fig. 1

SirT6 co-elutes with the H3K9me-specific HMTs Suv39h1 and G9a. a In vitro HMT assay of SirT6-HA using [³H]-labeled SAM and core histones as substrates. HA-affinity purification of extracts from 293F cells transfected with either empty vector (C) or SirT6-HA. As a loading control, a Coomassie blue-staining of the PVDF membrane is shown (CBB). b HMT assay performed as in a, but using, as substrate, purified recombinant proteins formed by GST fused to the N-terminal H3 histone tail (with the indicated mutations). GST-fused N-terminal H3 histone tails proteins were stained with CBB. c HA immunoprecipitation of extracts from 293F cells transfected with SirT6-HA together with each of the nuclear H3K9 HMTs. d Schematics of the different Myc-tagged constructs of Suv39h1 used in e. e HA immunoprecipitation of extracts from 293F cells transfected with SirT6-HA and the indicated Myc-tagged mutants of Suv39h1. f Immunofluorescence analysis of H3K9me3 and DAPI in wild type (wt), sirt1^−/−, and sirt6^−/− MEFs. A representative 5 μm scale bar is included in DAPI sirt1 wt image