Figure 1
From: Excess HB-EGF, which promotes VEGF signaling, leads to hydrocephalus
Effect of HB-EGF on endogenous VEGF in vitro.
(a) VEGF mRNA expression in mouse endothelial cell (CD31+) treated with 0, 10 and 100 ng/ml of recombinant human HB-EGF. (a’) A phase-contrast micrograph displaying endothelial cell reaching approximately 70% confluent state prior to HB-EGF treatment. (b) VEGF mRNA expression in mouse non-endothelial cell (CD31−) treated with 0, 10 and 100 ng/ml of recombinant human HB-EGF. Error bar represents the standard error (n = 4 for 1, 3, 6 and 12 h; n = 4 for 24 and 72 h data, respectively). (b’) A phase-contrast micrograph displaying non-endothelial cell reaching approximately 70% confluent state prior to HB-EGF treatment (c) Micrographs displaying CD31+ mouse brain endothelial cells cultured 3 day following magnetic isolation through the manual cell separation columns (MACS). (c’) The CD31 positive cells are clustered in the absence of αSMA positive vascular smooth muscle cell. The CD31 negative cell population is heterogenous, containing αSMA positive cells along with the cellular phenotype that are negative with anti-CD31 and negative with anti-αSMA stain (arrow). (c’) A representative micrograph demonstrating CD31 positive vascular endothelial cells on day 12 of the culture following MACS based isolation used in the exogenous HB-EGF experiments. Four independent replicate experiments from four C57BL6 mice (n = 4). Scale bars, 40 μm (c–c’) and 20 μm (c”) (d) Bar graph exhibiting VEGF protein levels in culture media of mouse endothelial cell treated at 0, 10 ng, 100 ng and 1 μg/ml of HB-EGF. Four replicate experiments (n = 4) in this figure (d). Single asterisk denotes a statistical significance by Dunnett’s post-hoc test with respect to the control after ANOVA (P < 0.05)
