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. 2008 Sep 8;182(5):979–991. doi: 10.1083/jcb.200712110

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© 2008 Ren 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 5. — Knockdown of IGFBP-5 suppresses IGF-II gene expression and decreases IGF-IR–mediated signaling activity. (A) C2C12 cells transfected with pSUPER or pSUPER-BP5 were induced to differentiate by switching to DM. RNA samples were prepared at the indicated time points and subjected to RT-PCR. (B) RNA samples described in A were analyzed by qRT-PCR. Data are expressed as relative value to those of the pSUPER transfected cells at 24 h after the induction of differentiation. Data shown are means ± SE of three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001 compared with the pSUPER-transfected control groups at the same time point. (C) Cells transfected with pSUPER or pSUPER-BP5 was induced to differentiate. 48 h later, cells were lysed and analyzed by Western immunoblot. The phospho-Akt/total Akt ratio was calculated by densitometry. Values are expressed as relative levels to pSUPER group. Data shown are means ± SE; n = 4; *, P < 0.05.