Figure 6

Lung tissue immunoblots. Pulmonary VEGF expression was higher in the bivalirudin group compared to controls (A,F) while there were no significant differences in the heparin and argatroban groups. Activation of VEGFR2 (phosphorylated/total receptor VEGFR2) was significantly lower in heparin-treated mice (G). There were no significant differences in the expression of the co-receptors neuropilin-1 (NRP-1) or neuropilin-2 (NRP-2) in then bivalirudin, argatroban, or heparin groups (B,H,I). Activation of the downstream proliferation marker ERK (phosphorylated/total receptor ERK) was not significantly different in the anticoagulated groups (C,J). Heparin-treated mice demonstrated increased activation of the downstream mediator Akt (phosphorylated/total Akt; D,K). Activation of an alternative pathway involving the epithelial growth factor receptor (phosphorylated/total EGFR) was lower in the argatroban and heparin groups, although not reaching statistical significance (E,L). Uncropped blots are provided in Supplementary Information Files 1,2. The β-actin for each membrane is displayed below the corresponding antibodies and was used to normalize the expression patterns. Each lane represents a sample from a different mouse. Statistical analysis of protein expression was done using analysis of variance (ANOVA), with Dunnett’s adjustment for multiple comparisons. Results are expressed as mean ± standard error. *P < 0.05.