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 inhibits LPS-induced myddosome formation. Murine bone marrow-derived DC (cultured for 4 d) conferred thalidomide (30 μM), PYR-41 (5 μM), or DMSO treatment prior to LPS (50 ng/ml) stimulation. The relocation of TLR4 (a) and MyD88 (b) was determined by confocal microscope by EEA1, TLR4, and MyD88 antibody staining. Nuclei were counterstained with DAPI (blue). Original magnification, ×600. The colocalized plots of TLR4 (a) or MyD88 (b) with EEA1 were counted and analyzed. (c–d) The effects of thalidomide and PYR-41 on TLR4 expression on DC were determined via flow cytometry. Numbers in histogram indicate MFI of analyzed population. The data were presented as the mean ± SEM, ,, and one-way ANOVA with Newman–Keuls post test.