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. 2002 Apr 1;157(1):45–62. doi: 10.1083/jcb.200112127

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Copyright © 2002, The Rockefeller University Press

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Figure 5. — The SNARE motif-related domain of p115 does not disrupt Giantin-p115-GM130 tethers. (A) RLGs were dissolved in Triton X-100 buffer and incubated for 1 h on ice in either the presence or absence of p115 (250 nM) plus or minus 50 μM CC1 or GM130 NT. Giantin was then immunoprecipitated, and coprecipitation of GM130 and p115 was determined by immunoblot. The input lane reflects 2% total p115 input and 10% total GM130 and Giantin input. (B) MGFs were incubated at 37°C for 1 h with p97/p47 plus or minus p115 (130 nM). 13 μM CC1 or GM130 NT was added to some reactions. Reactions were terminated by fixation, processed for EM, and the amount of relative cisternal regrowth was determined. 100% relative cisternal regrowth represents an increase from 25 to 75% of the total membrane present as cisternae. The percentage total membrane present as stacks was also determined. Values represent means ± SEM (n = 2).