Figures & data
Figure 1. Northern blot analysis for the expression of iNOS mRNA in human PMN. Cells were treated with 0.74 µg NDMA/µl for 2 h. Total RNA (10 µg/sample from 107 total cells) were then size-fractioned by agarose gel electrophoresis and transferred onto a positively-charged nylon membrane. Detection of iNOS mRNA or β-actin mRNA was performed using oligonucleotide probes labeled with biotin. Hybridization was followed by chemiluminescent detection of hybrids and analyzed using ImageJ software. Similar results were obtained from 10 independent experiments; one representative blot is shown.
Figure 2. Expression of iNOS, Fra-1, Fra-2, JunB, JunD, and FosB in PMN. (A) PMN were treated with or without SB203580 (40 µM), BIX02189 (30 µM), or SP600125 (40 µM) for 1 h before addition of NDMA (0.74 µg/µl). The cytoplasmic and nuclear fractions obtained from the cells were used to detect iNOS, Fra-1, Fra-2, JunB, JunD, and FosB protein levels by Western blot. The results shown are representative of five independent experiments. (B) Band intensity was quantified using ImageJ software and expressed in arbitrary units (AU). Data shown are mean (± SE) of five independent experiments. * Value significantly different between cells without and with NDMA (p < 0.05); #value significantly different between cells treated with NDMA but pre-incubated without or with the inhibitor (p < 0.05).
Figure 3. Concentrations of total NO from PMN. PMN were treated with or without SB203580 (40 µM), BIX02189 (30 µM), or SP600125 (40 µM) for 1 h before addition of NDMA (0.74 µg/µl). Two hours after addition of NDMA, the nitrite concentrations were measured as a marker of NO production. *Value significantly different between cells without and with NDMA (p < 0.05); #value significantly different between cells treated with NDMA but pre-incubated without or with the inhibitor (p < 0.05). Data are expressed as µM/106 cells (in 270 µl supernatant) and are shown as mean (± SE) of 20 experiments.
