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. Author manuscript; available in PMC: 2009 May 1.
Published in final edited form as: Dev Cell. 2008 May;14(5):661–673. doi: 10.1016/j.devcel.2008.02.004

Figure 2. SIRT1 Mediates the Effects of Glucose Restriction on Skeletal Myoblasts.

Figure 2

(A) Extracts of C2C12 cells cultured in DM with either 25mM or 5mM glucose and without or with NAM (5mM) were immunoblotted with the MHC, caveolin-3, and tubulin antibodies. (B) C2C12 cells were transduced with a retrovirus expressing a RNA hairpin directed against SIRT1 (shSIRT1) and cultured in DM with 25mM, 5mM, or 2.5mM glucose. Immunoblot was performed with antibodies against MHC, SIRT1, and tubulin. (C) MHC IF of cells expressing control or hSIRT1 cultured in DM with 25mM or 5mM glucose. (D) RT-qPCR of transcripts (embryonic myosin heavy chain Myh3, troponin C2, and SIRT1) derived from freshly isolated mouse primary skeletal myoblasts of 4 weeks-old wild-type (WT) or SIRT1+/− heterozygous mice cultured in DM with 25mM or 0.5mM glucose. Three independently obtained RNA samples were evaluated. The values of each transcript were corrected for those of GAPDH transcripts. Error bars represent standard deviations (p< 0.05). (E) MHC IF of WT and SIRT1+/− skeletal myoblasts cultured in DM with 25mM or 0.5mM glucose. (F) SIRT1 and tubulin immunoblot of extracts from mouse primary myoblasts obtained from either WT or SIRT1+/− heterozygous animals cultured in either 25mM or 0.5mM glucose.