Fig. 8
PSMC5 interacts with TLR4. A Colocalization of PSMC5 (red) and TLR4 (green), with DAPI nuclear counterstain in the BV2 microglia. Scale bar = 10 µm. B In silico modeling of PSMC5 interactions with TLR4. C Coimmunoprecipitation of endogenous PSMC5 and TLR4 in BV2 cells; precipitation was carried out with TLR4 antibody or IgG followed by WB analysis with antibodies against PSMC5 and TLR4. D GST-pull down to verify the computer molecular docking results. E PSMC5 expression after PSMC5 knockdown in the PSMC5 wild-type and PSMC5 mutants. F Western blot analysis of protein expression of the inflammatory proteins iNOS and COX-2, and MyD88-dependent signaling pathway components in the PSMC5 wild-type and mutants. G Results of the Griess assay showing NO content in the PSMC5 mutants. H ELISA analysis showing the PGE2 content in the PSMC5 mutants