PYR-41 and Thalidomide Impair Dendritic Cell Cross-Presentation by Inhibiting Myddosome Formation and Attenuating the Endosomal Recruitments of p97 and Sec61 via NF-κB Inactivation
Treatment with PYR-41 or thalidomide abrogates the endosomal recruitment of Sec61β. Murine bone marrow-derived DC (cultured for 4 d) conferred thalidomide (30 μM), PYR-41 (5 μM), or DMSO treatment prior to ovalbumin (50 μg/ml) or PBS pulse. The relocation of Sec61β (a) from endoplasmic reticulum to endosomes was determined by confocal microscope by Rab5, calnexin, and Sec61β antibody staining. The colocalized plots of Sec61β (b) with Rab5/calnexin were counted and analyzed. The expressions of Rab5/Sec61β and calnexin/Sec61β in the cells which are corresponding to the (a) colocalized cells were shown (c). Nuclei were counterstained with DAPI (blue). Original magnification, ×600. Data were presented as the mean ± SEM, , , and one-way ANOVA with Newman–Keuls post test. One representative from 3 independent experiments was shown. Rab5: early endosome marker; calnexin: endoplasmic reticulum marker.