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. 2011 Sep 1;22(17):3218–3230. doi: 10.1091/mbc.E10-11-0874

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© 2011 Eyster 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: — TSG101 knockdown reverses the effect of MARCH expression on CIE cargo proteins. HeLa cells were mock transfected (A, C) or transfected with an siRNA to TSG101 for 24 h (B, D). Cells were subsequently transfected with MARCH8-FLAG for 18 h (A–D). Total CD44 (A, B) or CD98 (C, D) distribution was detected with mouse antibodies and MARCH8-FLAG transfectants with a rabbit anti-FLAG antibody. Primary antibodies were detected with Alexa 488 (green) anti–mouse secondary and Alexa 594 anti–rabbit secondary, respectively, in mock (A, C) and TSG101 knockdown cells (B, D). (E) Western blot of mock and TSG101 knockdown lysates from replicate cultures. TSG101 protein levels (relative to actin) were reduced to 25% of control levels in siRNA-treated cells. Bars, 10 μm.