Figure 4.

Activation of autophagy by rapamycin reduces misfolded HLA-B27. (A) BM macrophages were incubated without or with IFNγ (50 ng/ml) for 6 hours, washed, and treated without or with rapamycin (25 μM) (Ra) for 2 hours. HC10 was used to immunoprecipitate unfolded/misfolded heavy chains, followed by immunoblotting with 3B10.7 (α-HLA). Co-precipitating BiP was measured using an aliquot of the immunoprecipitate run on a separate reducing gel. Immunoblots depict representative results. Quantitative results are mean ± SEM from six independent experiments, showing monomers, dimers, oligomers, and co-precipitating BiP. Inset shows longer exposure of oligomers. Dimers, oligomers, and BiP were quantitated only in IFNγ-treated samples. (*, p < 0.05)