Figure 2. DM mutant NOD mice weakly express A^g7/CLIP complexes and entirely lack mature compact A^g7 dimers.

(A) To evaluate occupancy by the Ii chain-derived CLIP peptide, DM mutant spleen cells expressing either the b or k haplotype, DM loss of function and wildtype NOD splenocytes were compared. Lysates were prepared from spleen cells pulsed for 40 mins with ³⁵S-methionine and chased for different times as indicated and extensively pre-cleared with Rabbit anti-mouse IgG (H+L) Abs before immunoprecipitation with beta chain-specific Rabbit Ab. Complexes were boiled for 10 mins and analyzed on 10% polyacrylamide gels. (B) Spleen cell lysates were boiled (B) or kept on ice (NB) and samples resolved on 10% gels under reducing conditions, transferred to nitrocellulose membranes, and Western blots probed with 10-2-16 or alpha chain-specific Rabbit Ab as indicated. (C) Western blot analysis of lysates prepared from LPS-stimulated BMDCs.