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Figure 2 | Journal of Neuroinflammation

Figure 2

From: Pro-inflammatory gene expression and neurotoxic effects of activated microglia are attenuated by absence of CCAAT/enhancer binding protein β

Figure 2

C/EBPβ expression in activated mixed glial cultures. Effect of 100 ng/mL LPS alone or in combination with 0.1 ng/mL IFNγ on C/EBPβ expression in secondary mixed glial cultures. A. C/EBPβ mRNA expression is upregulated in glial activation. Cultures were treated with LPS and LPS+IFNγ for 6 h and mRNA was analyzed by qPCR. Results are expressed as relative fold units (r.f.u.) of ΔΔCt values between C/EBPβ and actin + Rn18s as reference genes. One-way ANOVA followed by Dunnett's test is applied. *p < 0.05; **p < 0.01 compared to control. (n = 3). B. LPS and LPS + IFNγ (24 h) increase nuclear C/EBPβ immunostaining (red) in secondary mixed glial cultures. In the right upper corner, a detail shows overlapping between Hoechst 33258 nuclear staining and C/EBPβ. Images are representative of 5 independent experiments. Bar = 50 μm. C. A western blot shows levels of C/EBPβ in secondary mixed glial cultures treated with LPS or LPS + IFNγ for 24 h. The C/EBPβ isoforms are identified as Full/LAP and LIP. β-Actin is used for normalization. This experiment was done 4 times with similar results. D. Full/LAP (grey bars) upregulation after LPS and LPS+IFNγ is statistically significant compared to control. LIP (dashed bars) upregulation is statistically significant only for LPS treatment. One-way ANOVA, followed by Dunnett's test is applied. *p < 0.05; **p < 0.01 compared to respective control. (n = 4-5)

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