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. 2013 Sep 26;9(9):e1003747. doi: 10.1371/journal.pgen.1003747

Figure 2. AUF-1 enhances IL-8 expression by halting IL-8 mRNA degradation.

Figure 2

a) AUF-1 (approx. 37 kD) was knocked down in NCI-H292 cells with si-AUF-1. Scrambled si-RNA (si-con) served as a control for specificity and β-actin (42 kD) was determined to verify equal protein loading. b). IL-8 released over 24 h in supernatant from NCI-H292 cells, non-transfected (nt) or transfected with si-con or si-AUF-1 and either non-stimulated (ns) or stimulated with IL-17, TNF-α or TNF-α plus IL-17. Data are shown as mean ± SD. ***P<0.001 (2-way ANOVA, Bonferroni's multiple comparison post-test). c). Q-PCR analysis of IL-8 mRNA was done for non-transfected or transfected cells with si-con or si-AUF-1 at 2 h after stimulation with TNF-α or TNF-α plus IL-17, or left unstimulated. **P<0.01 (2-way ANOVA, Bonferroni's multiple comparison post-test). mRNA results are presented relative to that of unstimulated, non-transfected cells d). Non-transfected, si-con or si-AUF-1 transfected cells were left unstimulated or stimulated with TNF-α or TNF-α plus IL-17 for 2 h. RNA was isolated from cells at 0, 30 and 60 mins after blocking gene transcription by actinomycin D. % of IL-8 mRNA remaining was calculated on basis of q-PCR. Representative experiments are shown for 3 experiments.