Research Article
Targeted Delivery of the HLA-B27-Binding Peptide into the Endoplasmic Reticulum Suppresses the IL-23/IL-17 Axis of Immune Cells in Spondylarthritis
Figure 2
The effect of treatment with THUA or THUNP on the (B27-HC)2 production of C1R-B27:04 cells. After C1R-B27:04 cells were treated with each recombinant protein for the indicated time, membrane proteins were extracted. 50 μg of extract was analyzed by nonreducing SDS-PAGE (10%) and immunoblotted with a BH2 monoclonal antibody and anti-transferrin (Tf) receptor antibody. Tf receptor acts as an internal control. (a) THUA treatment for 12 h, but not THU or HUA treatment, significantly decreased the levels of (B27-HC)2. (b) The ratio of (B27-HC)2/Tf receptor averaged from three independent experiments in (a) is plotted against the time of THU, HUA, or THUA treatment (mean ± SD, ). (c) Treatment of C1R-B27:04 cells with THU, HUA, HUNP, THUA, THUNP, or TAP-1 knockdown does not affect the expression levels of B27-HC. C1R-B27:04 cells were treated with 20 μg of THU, HUA, HUNP, THUA, or THUNP for 24 h. The membrane proteins were extracted. An aliquot (50 μg) of the extracted protein was analyzed by reducing SDS-PAGE and immunoblotted with the BH2 monoclonal antibody. (d) Treatment with the THUNP reduces the formation of (B27-HC)2. (e) The ratio of (B27-HC)2/Tf receptor averaged from three independent experiments in (d) is plotted against the time of THU, HUNP, or THUNP treatment (mean ± SD, ). (f) TAP1-knockdown suppresses the THUNP-induced reduction of (B27-HC)2. (g) The ratio of (B27-HC)2/Tf receptor averaged from three independent experiments in (f) is plotted against the time of THUNP treatment (mean ± SD, ). values were obtained by the Mann–Whitney U test.
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