Journal: PLoS Neglected Tropical Diseases

Article Title: Live Brugia malayi Microfilariae Inhibit Transendothelial Migration of Neutrophils and Monocytes

doi: 10.1371/journal.pntd.0001914

Figure Lengend Snippet: HUVEC or HLMVEC were cultured with or without live untreated B. malayi Mf or Wolbachia -depleted Mf for 24 hours prior to isolation of total EC RNA. An oligo microarray was used to analyse mRNA expression of various angiogenesis mediators (a). mRNA expression was compared between HUVEC in the absence of Mf vs. presence of intact Mf and HUVEC in the absence of Mf vs. presence of Wolbachia -depleted Mf. HUVEC and HLMVEC mRNA expression of (b) Ang-2, (c) BAI-1, (d) TNFSF15 and (e) COX-2 was also analysed by real time qRT-PCR. Data are shown as the mean of two independent experiments (oligo microarray) or mean and standard deviation of three independent experiments (qRT-PCR). Students t-test was used to analyse statistical significance. (f) HO-1 protein expression in HLMVEC is enhanced by live Mf presence. HLMVEC were left unstimulated or were stimulated with IFN-γ for 24 or 48 hours, prior to co-culture with live Mf for a further 6, 12 or 24 hours. HLMVEC cells were lysed and the cell lysate was analysed for protein levels of HO-1 by western blotting.

Article Snippet: Blots were incubated with mouse anti-human heme oxygenase-1 (HO-1) (BD transduction Laboratories) or mouse anti-human β-actin antibodies.

Techniques: Cell Culture, Isolation, Microarray, Expressing, Quantitative RT-PCR, Standard Deviation, Co-Culture Assay, Western Blot