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 abolishes the endosomal recruitment of p97. Murine bone marrow-derived DC (cultured for 4 d) firstly conferred thalidomide (30 μM), PYR-41 (5 μM), or DMSO treatment prior to ovalbumin (50 μg/ml) or PBS pulse. The relocation of p97 from endoplasmic reticulum to endosomes (a) was determined by confocal microscope by Rab5, calnexin, and p97 antibody staining. The colocalized plots of p97 with Rab5 and calnexin (b) were counted and analyzed. The expressions of Rab5/p97 and calnexin/p97 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.