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. 2012 Mar 15;23(6):1095–1103. doi: 10.1091/mbc.E11-08-0668

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© 2012 Urbé 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 8: — USP21 depletion interferes with NGF-induced neurite outgrowth in PC12 cells. PC12 cells were transfected with pRFP-C-RS vector expressing either nontargeting shRNA (NC15, Control) or HuSH-29 shRNA targeting rat USP21 (shUSP21#1–4). Transfected cells coexpress RFP. Neurite outgrowth was induced by application of 100 ng/ml NGF 48 h after transfection, and 3 d later cells were fixed and counterstained with Alexa Fluor 488–labeled phalloidin to visualize nontransfected cells. Cells expressing RFP were scored as differentiated if one or more neurites had reached a length larger than the diameter of the cell body (30–48% of control cells, compared with 4–15% in USP21-depleted cells). The data are derived from three independent experiments, in each of which between 130 and 350 cells were scored for each condition. Error bars, ±SD.