Fig. 1

Effect of inflammation on ANG2 expression and release. a HMVECs were incubated with LPS (1 μg/ml), thrombin (1 U/ml), histamine (100 μM), or Phorbol 12-myristate 13-acetate (PMA; 50 ng/ml), respectively. The upper panel shows ANG2 release into cell culture supernatant after incubation for 1 h, measured by ELISA. The lower panels show expression of ANG2 (left) and IL-8 (right), respectively, quantified by RT-PCR after 3 h of stimulation. Relative target gene expression was normalized to 18 s RNA and is shown as fold change of expression of the unstimulated control. Mean from biological triplicates ±SD. b Female C57BL/6 mice (10–12 weeks old) were challenged by intratracheal instillation of LPS (5 μg/g bodyweight), and 24 or 72 h thereafter, animals were sacrificed and the lungs were harvested. The upper panel shows on the left a representative western blot analysis of lung ANG2 protein content (molecular weight 140 kDa), together with the corresponding histone band (cropped from image of Ponceau staining of the same nitrocellulose membrane). On the right, the results from the densidometric analyses are given, normalized to the histone band (mean ± SD from three blots with identical results). The lower panels show ANG2 and TIE2 expression, quantified by RT-PCR. Relative target gene expression was normalized to 18 s RNA and is shown as fold change of expression of the PBS control (represented by the dashed line). Mean ± SD, n = 7. ALI = Acute Lung Injury. Mann-Whitney U test, *p < 0.05 (vs. unstimulated or PBS control, respectively)