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. 2009 Jun 12;297(2):L388–L400. doi: 10.1152/ajplung.90373.2008

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Fig. 5. — A: effect of inhibitor of JNK on activation of JNK in A549 cells treated with Ox-AT. Ox-AT treatment increased level of activated JNK but not total JNK significantly compared with PBS (*P < 0.001, P = 0.7, respectively). SP-600125 significantly inhibited this effect of Ox-AT on activation of JNK (**P = 0.016). SP-600125 did not have any significant effect on total level of JNK in cells of both PBS group and Ox-AT (P = 0.109 and 0.171, respectively). Data are means (SE) from 4 separate experiments, *^, **P < 0.05. B: Western blot analysis of NHBE cell lysate for phosphorylated JNK (phospho-JNK). Western blot analysis shows that p54 JNK isoform was expressed highly in Ox-AT (0.3 mg/ml)-treated cells compared with PBS-treated cells, which was further verified by densitometric analysis of each band using ImageJ software. Densitometric results show level of p54 phospho-JNK is significantly higher and peaking at 1 h in Ox-AT-treated cells compared with PBS-treated cells (0.5 h P < 0.001; 1 h P < 0.001; 2 h P = 0.043). Lane 1, positive control A431 cell lysate; lane 2, 0.5 h PBS; lane 3, 1 h PBS; lane 4, 2 h PBS; lane 5, 0.5 h Ox-AT; lane 6, 1 h Ox-AT; lane 7, 2 h Ox-AT. Gel is representative of 3 independent experiments. Data are means (SE) of 3 independent experiments, *P < 0.05. C: Western blot analysis of NHBE cell lysate for total JNK. Western blot analysis for total JNK shows total (phosphorylated and nonphosphorylated) JNK ranging in size between 46 and 54 kDa. Densitometric analysis of both bands from each sample (expressed cumulatively) using ImageJ software demonstrated that total JNK was expressed in comparable levels in both Ox-AT (0.3 mg/ml)- and PBS-treated cells. Densitometric analysis of bands showed there was no significant difference between Ox-AT- and PBS-treated cells at 0.5, 1, or 2 h. Lane 1, positive control A431 cell lysate; lane 2, 0.5 h PBS; lane 3, 1 h PBS; lane 4, 2 h PBS; lane 5, 0.5 h Ox-AT; lane 6, 1 h Ox-AT; lane 7, 2 h Ox-AT. Gel is representative from 3 independent experiments. Data are means (SE) of 3 independent experiments.