Figure 9.

TNF regulates Angpt2 expression in vitro. a, b, Quantification of Angpt2 mRNA by real-time PCR in bEnd.3 cerebral endothelial cells or primary astrocytes cultured for 6 h in the presence of LPS (1 μg/ml), TNF (10 or 100 ng/ml), and/or anti-TNF antibody (10 μg/ml). *Significantly different from control [ANOVA, p < 0.0001 (a) or = 0.0072 (b)]. Error bars indicate SEM. c, Confirmation of the presence of Angpt2 mRNA in bEnd.3 cells and astrocytes, cultured in normal conditions, by standard PCR and agarose gel electrophoresis. Right column, DNA ladder. d, Quantification of TNF mRNA by real-time PCR in bEnd.3 cells and astrocytes cultured for 6 h with or without LPS. *Significantly different from PBS (Student's t test, p < 0.0001). Error bars indicate SEM. e, Confocal image of astrocytes in primary culture immunostained for GFAP (red). Blue, DAPI-stained nuclei. Scale bar, 20 μm.