Fig. 2. Induction of pro-inflammatory mediators is attenuated in microglial primary cultures from C/EBPβ-deficient mice. (A) Western blots showing a reduction in the expression of IL1β and COX2 in C/EBPβ^–/– microglial cultures after treatment with LPS or KA for 24 hours, compared with cultures established from C/EBPβ^+/+ mice. (B,C) Primary microglial cultures prepared from C/EBPβ^+/+ or C/EBPβ^–/– mice were treated as in A, and the expression of C/EBPβ (green), IL1β (blue in B) and COX2 (blue in C) was evaluated using immunofluorescence techniques, and confocal microscopy using specific antibodies as described in the Materials and Methods. The last column of panels in each set shows colocalization of C/EBPβ with IL1β (B) or COX2 (C). The microglial marker CD11b is shown in red. Representative results from three experiments are shown. Scale bar, 10 µm. B, basal; KA, kainic acid; LPS, lipopolysaccharide.