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. 2008 Aug 11;182(3):497–507. doi: 10.1083/jcb.200801119

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© 2008 Kang et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 7. — Overexpression of Cdc42GAP or BPGAP reduces myogenic differentiation. (A) Lysates of C2C12 cells stably transfected with a control expression vector lacking a cDNA (−) or with expression vectors harboring flag-tagged Cdc42GAP or BPGAP cDNAs as indicated (+) were Western blotted with flag epitope or Cdc42GAP antibodies. (B) Levels of GTP-bound and total Cdc42 in C2C12/Cdc42GAP, C2C12/BPGAP, or vector control (−) cells in GM or DM were assessed as described in Fig. 6 A. (C) Photomicrographs of C2C12/Cdc42GAP, C2C12/BPGAP, and vector control cells that were cultured in DM, fixed, and stained with an antibody to MHC. Bar, 0.1 mm. (D) Quantification of myotube formation. Values represent means of triplicate determinations ±1 SD. The experiment was repeated three times with similar results. Asterisks indicate difference from control, P < 0.01. A level of myotube formation by control cells (∼80% nuclei in MHC⁺ cells) was selected so as to permit visualization of diminished differentiation by Cdc42GAP proteins. (E) Western blot analysis of muscle-specific proteins by C2C12 cell transfectants cultured in GM (G) or in DM for the indicated times.