Fig. 1

Elevated CD40 expression by LPS or IFNγ in the BV-2 and primary microglia cells is regulated by cPLA₂α. A representative double-immunofluorescence staining of cPLA₂α (green) and CD40 (red) in unstimulated or stimulated microglia by LPS or IFNγ in the absence or presence of AS or sense (SE). DAPI staining shows cell nuclei. The BV-2 cells were treated with (a) 50 ng/ml LPS or (b) 10 ng/ml IFNγ for 24 h. The mouse primary microglia cells were treated with (e) 50 ng/ml LPS or (f) 25 ng/ml IFNγ for 48 h. Scale bars = 50 μm. Four micrometer AS or the corresponding sense (SE) were added to the cultures 24 h before addition of the stimuli. The intensity of CD40 or cPLA₂α were quantitated for the cell and expressed in the bar graph as arbitrary units. A representaive immunoblot analysis of cPLA₂α and CD40 for the BV-2 microglia cells (c, d) and primary mouse microglia (g, h) treated as (a), (b), (e), and (f). The intensity of each cPLA₂α or CD40 band after quantification by densitometry was divided by the intensity of each calreticulin (Calre) band and expressed as arbitrary units. The bar graphs are the mean ± SE from three independent experiments. (***p < 0.0001)