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. 2014 Jun 10;9(6):e98459. doi: 10.1371/journal.pone.0098459

Figure 5. TNFα decreases cell NO, and reversal of this effect by adenosine is eNOS-dependent.

Figure 5

HMEC were plated in 6-well plates, then treated for 48 h as described for Fig. 4. Prior to treatment, some cells were transfected with morpholino-antisense or control oligos. Cells were washed free of media then loaded in the dark for 30 min with DAF-FM diacetate (5 µM) in HBSS+10 mM HEPES. Cells were further washed free of DAF-FM-containing buffer, and incubated a further 15 min. They were then gently scraped form the plate and separate aliquots were taken for assay of fluorescence and protein content. All fluorescence measurements were normalized to protein concentration, and the percent change in cellular fluorescence was calculated. Values are means ± S.E.M. for 4 separate experiments of 3 replicates each for each experimental condition except NOS3 (eNOS antisense) + detaNO for which data are for 3 separate experiments. Differing letters denote significant, between-group differences, p<0.05.