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. Author manuscript; available in PMC: 2015 Mar 18.
Published in final edited form as: FEBS Lett. 2014 Jan 31;588(6):906–914. doi: 10.1016/j.febslet.2014.01.033

Figure 1. Pro-inflammatory cytokines reduce S100A1 expression and cause dysfunction in ECs.

Figure 1

A) Primary HMVECs were incubated with Ang II (10 nmol/L), Et-1 (10 nmol/L), TNFα (50 ng/ml) or CoCl2 (250 μmol/L) for 24h before extract preparation and SDS-PAGE analysis for ICAM and S100A1. Representative image is shown. Experiment was repeated twice more, for quantification see Figure S1. B) HMVECs were incubated with cytokines for 24h, then treated ± VEGF (50 ng/ml) as indicated, for a further 24h. eNOS produced nitric oxide levels in the supernatant were measured using a nitrate/nitrite assay. Experiment was done 3 times, each at least in duplicate. C) HMVECs were infected with recombinant adenovirus expression S100A1 (or ctr. Adenovirus) and treated for 24h with Ang II, Et-1 or TNF as indicated. Cells were then stimulated, or not, with VEGF (50 ng/ml) for 24h before assaying supernatant nitrate/nitrite levels. *, P<0.05 vs untreated, no VEGF. # P<0.05 vs untreated, no VEGF; ✠, P<0.02 vs untreated + VEGF. Experiment was performed 3 times, each at least in duplicate.