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. 2014 Jul 9;111(29):10684–10689. doi: 10.1073/pnas.1411026111

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Fig. 4. — The interaction between FoxO1 and SIRT6 promotes FoxO1 nuclear exclusion. (A and B) Analysis of endogenous FoxO1 interactions with SIRT1, SIRT6, or HDAC1 in HCT116^(+/+) cells and HCT116^(−/−)cells. Anti-FoxO1 immunoprecipitates of nuclear extracts were probed with the indicated antibodies. (C and D) Analysis of recombinant FoxO1 interactions with SIRT1 and SIRT6 in the nuclear extracts of HCT116^(+/+) and HCT116^(−/−) cells. GST pull-down assays with bound proteins probed with the indicated antibodies. (E and F) Effects of SIRT6 knockdown on nuclear exclusion of FoxO1 following ectopic p53 expression in HCT116^(−/−) cells or adriamycin-induced p53 expression in HCT116^(+/+) cells. Nuclear extracts from CTR or SIRT6 RNAi-treated cells were subjected to immunoblotting with the indicated antibodies. (G and H) Effects of SIRT1 or HDAC1 knockdown on nuclear exclusion of FoxO1 following ectopic p53 expression in HCT116^(+/+) cells. Nuclear extracts from CTR and SIRT1 RNAi- and HDAC1 RNAi-treated cells were subjected to immunoblotting with the indicated antibodies.