Figure 5.

PAP suppressed LPS-induced microglial activation by NF-κB signaling pathway.

Notes: Cells were seeded on poly-L-lysine coated dishes and pretreated with PAP (10 μg/mL) for 4 hours. Then, LPS was added to the medium and incubated for another 1 hour. For immunofluorescence analysis, cells were fixed and incubated with primary/secondary antibodies. For Western blotting, cells were lysed and the total/nuclear protein was extracted for further analysis. A significant increase in nuclear translocation of P65 and remarkable upregulation of intranuclear P65 were noticed after 1 hour of LPS application, but this effect was partly abolished by 4 hours of PAP pretreatment (A and B). Consistent with the changes of P65, LPS stimulation increased phosphorylation of IKK in BV2 cells, but it was partly blocked by PAP pretreatment (C and D).

Abbreviations: PAP, papaverine; LPS, lipopolysaccharide; CON, control; DAPI, diamidino-phenyl-indole.