Additional file 2.

Efficiency of EC transfection with specific miRNA inhibitors and precursors. DIC (A) and immunofluorescence (B), immunostaining with anti-CD31 (green), confocal images of the ECs transfected with Cy-3-conjugated control miRNA inhibitor, depicted in red (24 hours post-transfection). C and D: Transfected ECs were plated onto Matrigel matrix for 24 hours and subjected to immunofluorescence staining with anti-CD31 antibody (green). Orthogonal image projection demonstrates the intracellular localization of the Cy-3-conjugated control (red). Bars: 20 μm on B; 10 μm on C. E and F: Efficiency of the miRNA inhibition and overexpression. ECs were transfected with pre-miR miRNA precursors (E) and anti-miR inhibitors (F) specific for miR155 and miR-100. At 24 hours after transfection, Q-RT PCR was performed using TaqMan preformulated primer sets specific for these miRNAs. The obtained raw data were analyzed using the SDS2.4 and RQ Manager1.2.1 software that converted the Ct values into fold-change values using the snoRNA202 as the sample normalizing RNA. miRNA expression fold changes (specific anti/pre-miR-transfected vs control anti/pre-miR-transfected cells) are displayed graphically.

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Roitbak et al. Vascular Cell 2011 3:25   doi:10.1186/2045-824X-3-25