Figure 6. Effects of puromycin on the recovery of cell surface MHC class I-peptide complexes.

A–E. 293-K^b cells expressing the TRx9 reporter were stripped of cell surface MHC I peptides as in Fig. 5. Recovery of cell surface MHC class I-peptide complexes was conducted in the presence of varying concentrations of puro from 0 to 180 minutes. Flow cytometry was used to measure reporter eGFP fluorescence (A), as well as cell surface K^b-SIINFEKL complexes (B) and total cell surface K^b (D). To quantitate differences in the kinetics of MHC class I-peptide complex recovery, we normalized the MFI values of puro-treated cells to untreated cells for K^b-SIINFEKL (C) and total K^b (E) as described in [4] (n = 5; mean ± s.e.m.) For C and E, * p<0.05 compared to untreated samples. F. MHC class I-peptide complex recovery after 1 hour in the presence of varying concentrations of puromycin. MFI values are normalized to untreated cells (n = 3; mean ± s.e.m.) G. MHC class I-peptide complex recovery after 4 hours in the presence of varying concentrations of puromycin. MFI values are normalized to untreated cells (n = 3; mean ± s.e.m.).