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. 2006 Dec 11;401(Pt 1):121–127. doi: 10.1042/BJ20060847

Figure 4. HLA-A2 ICD is produced under cell-free and in vivo conditions.

Figure 4

(a) Cell-free generation of a 12 kDa HLA-A2 ICD. Partially purified membrane fractions of CHO-HLA-A2 were subjected to a γ-secretase cleavage assay [8]. Membranes were incubated with protease inhibitors at 0 and 37 °C in the presence or absence of DAPT. Samples incubated at 37 °C generate a HLA-A2 ICD fragment. Blots were stained with an anti-V5 antibody. (b) In vivo generation of HLA-A2 ICD in CHO-HLA-A2 cells after incubation with a proteasomal inhibitor lactacystin. HLA-A2-ICD accumulation was blocked upon inhibition of PS1/γ-secretase activity with DAPT and L-685,458 respectively. The asterisk indicates an unidentified band that is not seen with PS1/γ-secretase inhibitor treatments.