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. 2010 May 31;189(5):829–841. doi: 10.1083/jcb.200912105

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© 2010 Pearse et al.

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Figure 2. — Reglucosylation supports persistent lectin chaperone association. (A) MI8-5 cells were pulse labeled for 1 h in the presence of 0.5 mM DNJ. Lysates were subjected to GST-calreticulin (CRT) pull-down (lanes 2 and 3) or immunoprecipitated with calnexin (CNX; lanes 4 and 5) or calreticulin (lanes 6 and 7) antisera. Some samples (+) were sequentially precipitated with saposin D antisera (αSapD). (B) Wild-type (WT) and ugt1^−/− cells were pulse labeled for 15 min and chased for the indicated times. Monoglucosylated prosaposin (Psap) was isolated by GST-calreticulin pull-down. DNJ was added at the 0.5-h time point until the completion of the assay for the indicated lanes. Samples were resolved via 7.5% SDS-PAGE. (A and B) Molecular mass is indicated in kilodaltons. (C) Quantifications of B using n = 3 and mean ± SD (error bars).