Fig. 4

Inhibition of LPS-induced p65 translocation by synaptamide. BV2 murine microglia cells were incubated with LPS (100 ng/mL) with or without synaptamide (10 nM) and OEA (10 nM) for 30 min. Adenylyl cyclase inhibitor (SQ 22536, 10 μM) and PKA inhibitor (H-89, 10 μM) were added 30 min prior to LPS/synaptamide treatment. Translocation of p65 was determined by immunocytochemistry (red, Alexa Fluor 588; blue, DAPI) (a). Nuclear levels of p65 were determined by immunoblot analysis. α-Tublin and lamin A/C were used as internal controls for cytosolic and nuclear fractions (b). Densitometric analysis of the western blots is represented as the mean band density normalized to lamin A/C. Values are the means ± SEM (n = 3). Means designated with the same letter are not significantly different. In A, the fluorescence intensity profile across a transverse section of one cell indicated in the second column is shown in the fourth column