FIG. 5.

Information in the final 12 amino acids of mK3 is critical for TAP/tapasin interaction. (A) Schematic depiction of the C-terminal domain (CT) mutants of mK3. NT, N-terminal domain. In the mK3/kK5intCT construct, mK3 residues 140 to 189 were replaced by kK5 residues 147 to 194. (B) Each mutant was introduced into B6/WT3 cells. Flow cytometric analysis of K^b versus GFP fluorescence is shown. The numbers in each plot represent the ratio of K^b staining intensities between GFP⁺ and GFP⁻ fractions. Analysis of D^b expression yielded very similar results (data not shown). (C) The GFP⁺ cells from each line shown in panel B were enriched by sorting. Lysates from these cells were blotted with the indicated antibodies following culture for 24 h with or without 100 U of IFN-γ/ml. GFP staining levels serve as a loading control as well as an indicator of the effect of IFN-γ on pMIG expression. Bands corresponding to mK3CT190 and mK3CT199 are indicated by arrowheads.