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. 2003 Sep 5;100(19):10794–10799. doi: 10.1073/pnas.1934713100

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Copyright © 2003, The National Academy of Sciences

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Fig. 1. — Generation of SIRT1-deficient mice. (A) Western analysis of SIRT1 and tubulin expression in WT adult mouse tissues. (B) A schematic diagram of the germ-line SIRT1 locus, KOII targeting vector, and SIRT1^Δ^ex4 allele. SIRT1 exons comprising the conserved catalytic domain are dark gray. The relative locations of the 5′, 3′, and internal KOII probes are indicated. Restriction sites: BgI, BglI; N, NotI; S, SalI; H, HindIII; E, EcoRI; Bg, BglII; B, BamHI; Pac, PacI; Swa, SwaI. (C) Southern blot of SIRT1⁺^/^Δ^ex4, SIRT1⁺^/NeoR, and SIRT1^Δ^ex4/^Δ^ex4 BglII-digested DNA with the internal KOII probe. (D) Western blot of SIRT1⁺^/⁺, SIRT1⁺^/^Δ^ex4, and SIRT1^Δ^ex4/^Δ^ex4 MEFs. SIRT1 and SIRT1^Δ^ex4 proteins are indicated. (E) A schematic diagram of the germ-line SIRT1 locus, SKO targeting vector, and SIRT1^Δ allele, which are depicted as in A. (F) Southern blot of SIRT1⁺^/^Δ^neo and SIRT1⁺^/^Δ BglI/SwaI-digested DNA with the 5′SKO probe. (G) Western blot of SIRT1⁺^/⁺, SIRT1⁺^/^Δ, and SIRT1^Δ^/^Δ MEFs with anti-SIRT1 antibodies. Arrows indicate crossreacting bands that control for total protein.