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. 2006 Feb 27;172(5):719–731. doi: 10.1083/jcb.200510065

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Copyright © 2006, The Rockefeller University Press

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.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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Figure 9. — IRS-2 activation-mediated autophagy in a conditional neuronal cell line. (a) N2a stable cell lines inducibly expressing exon1htt-25QmCFP and -103QmCFP. Nuclei are stained with Hoechst 33342. (b) Clearance of accumulated protein occurred in 6 d. Data represented as mean ± SEM. (c) IRS-2 and Beclin 1 are required for IGF-1–mediated clearance. ANOVA revealed a significant effect across groups (F_(4,43) = 5.793; P = 0.0008) Fisher's protected least significant difference test for “%Control” revealed a significant difference between control and dox-treated cells (P = 0.0006) and control and IGF-1–treated cells (P = 0.0179). Coadministration of siRNA against beclin1 (P = 0.9611) or IRS-2 (P = 0.4712) with IGF-1 eliminated clearance. Data represented as mean ± SEM. Asterisks indicate statistical significance, as indicated. (d) Quantification of the mCFP-positive inclusions found in LC3-positive autophagosomes. ANOVA revealed a significant effect across group (F_(3,16) = 3.873; P = 0.0294). Fisher's protected least significant difference test for “% cells with LC3-positive inclusions” revealed a significant difference between control and IGF1 (P = 0.0097), but no difference with siIRS-2 (P = 0.7881) or siBeclin1 (P = 0.8372). Data represented as mean ± SEM. Asterisks indicate statistical significance, as indicated.