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. 2005 Aug 15;202(4):505–516. doi: 10.1084/jem.20050575

Figure 3.

Figure 3.

XBP-1 does not affect the synthesis of κ light chain or synthesis and trafficking of class I and class II MHC. Cells stimulated for 3 d with CpG were pulse-labeled with [35S]methionine for 30 min and chased for up to 3 h. Cells were lysed in lysis buffer containing 0.5% NP-40. Under these conditions, MHC class I and class II complexes are preserved. (A) 90% of input were immunoprecipitated with P8 to recover H-2Kb. Class II I-A were immunoprecipitated with JV2 antibody. Immunoprecipitates were analyzed by SDS-PAGE (10%). Note the nonspecific recovery of μ chains only from WT cells. (B) 10% of input was immunoprecipitated with anti-μ antibodies. (C) Cells were stimulated for 3 d with CpG or LPS. 106 cells were pulse-labeled with [35S]methionine for 10, 20 or 30 min. Cells were lysed in 1% SDS, and the lysate was diluted to 0.07% SDS with NP-40 lysis mix. 70% of lysis mix was immunoprecipitated with P8, 20% was immunoprecipitated with anti-μ, and 10% was immunoprecipitated with anti-κ. Immunoprecipitates were analyzed by SDS-PAGE (12%). (D) Autoradiograms were quantified by phosphoimager. Expression levels of μ and κ were normalized to the corresponding levels of H-2Kb.