Mammalian Vectors for FBXO7 and SIRT7

Mammalian expression vectors for FLAG-tagged human wildtype FBXO7 isoform-1 and its F-box deleting mutant were provided by Dr H.J. Kuiken (The Netherlands Cancer Institute). The mammalian expression vector for V5/His-tagged human wildtype SIRT7 was provided by Dr K.Y. Choi (Pohang University of Science and Technology). Plasmids encoding FLAG-tagged SIRT2, SIRT3, SIRT4, and SIRT7 were kindly provided by H.S. Kim (Ewha Womans University). Plasmids encoding FLAG-tagged SIRT1, SIRT5, and SIRT6 were provided by Y.J. Oh (Yonsei University). All plasmid sequences were verified using DNA sequencing (Bionics). siRNAs for FBXO7, SIRT7, and CUL1, and control scrambled siRNA (catalog no.: 51-01-14-04) were designed and synthesized by Integrated DNA Technologies (Hanam-si). FBXO7-specific siRNA duplex sequences were 5′-UUGGUUCUCCUCUAGAUUGAAGU(dTdT)-3′ (sense) and 5′-ACUUCAAUCUAGAGGAGAACCAA(dTdT)-3′ (antisense). SIRT7-specific siRNA duplex sequences were 5′-GUGUGAACUUUAUAGAAU(dTdT)-3′ (sense) and 5′-AGAGAGGAUUCUAUAAAG(dTdT)-3′ (antisense). The CUL1-specific siRNA duplex sequences were 5′-CAGGUUUACCUUCAUGAAAGCACAC(dTdT)-3′ (sense) and 5′-GUGUGCUUUCAUGAAGGUAAACCUGAA(dTdT)-3′ (antisense).