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. 2015 Oct 1;26(19):3390–3400. doi: 10.1091/mbc.E15-05-0321

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© 2015 Pisoni 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 4: — Client-mediated association between TMX1 and CNX. (A) MEFs were cotransfected with an empty vector (EV, lanes 1 and 2), an empty vector and HA-tagged TMX1_C/A (3 and 4), an empty vector and BACE501 (5 and 6), or BACE501 and HA-tagged TMX1_C/A (7 and 8). Cells were incubated for 10 h in the absence (–) or presence (+) of CST (1 mM). The expression level of BACE501 was checked upon WB of the immunoisolated ectopic protein. (B) Same as A, but endogenous CNX with associated proteins was immunoisolated from cell lysates. Immunocomplexes were analyzed under reducing conditions. Ectopically expressed BACE501 and TMX1_C/A were revealed with an anti-BACE and an anti-HA antibody, respectively. (C) Same as A, in cells treated with CST, cycloheximide (Chx), or tunicamycin (Tun) for 3 h. (D) Same as B for cells treated with CST, Chx, or Tun. G, mature Golgi form of BACE501; E, immature ER form; D, deglycosylated form.