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. 2011 Aug 15;22(16):2797–2809. doi: 10.1091/mbc.E11-02-0137

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© 2011 Matsumura et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 6: — Hsc70 is required for CFTR dislocation. (A) Microsomes containing newly synthesized CFTR were subjected to in vitro degradation in the presence or absence of CBag. At indicated times, aliquots were separated into pellet (membrane) and supernatant (cytosol) fractions. Graph shows the total amount of CFTR released into the cytosol compared with percentage of TCA-soluble CFTR at each time point (mean ± SEM, n = 4). Bottom panels show phosphorimage of membrane and cytosol fractions. (B) CFTR degradation was carried out as in (A), but in the presence of both MG132 and CBag. Graph shows TCA-soluble and total CFTR released from membranes (mean ± SEM, n = 4), whereas gels show SDS–PAGE and phosphorimaging. In the presence of MG132, CFTR is released from membranes as both a HMW species and partially degraded fragments. In the presence of CBag, both of these forms were barely detected.