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Fig. 6 | Cell & Bioscience

Fig. 6

From: Dimerized translationally controlled tumor protein increases interleukin-8 expression through MAPK and NF-κB pathways in a human bronchial epithelial cell line

Fig. 6

Role of p38 MAPK on IL-8 mRNA stability in BEAS-2B cells. a BEAS-2B cells were pretreated with vehicle or SB203580 (10 μM) for 30 min, and stimulated with dTCTP (10 μg/ml) for 1 h. Cell lysates were immunoblotted with the indicated antibodies. b Nuclear extracts were obtained from dTCTP (10 μg/ml, 1 h) stimulated cells pretreated with SB203580 (10 μM) for 30 min. Nuclear extracts without stimulation were also prepared as a negative control. They were subjected to EMSA analysis using biotinylated probes and the specificity of the binding reaction was confirmed using 10-molar excess of unlabeled oligos. The results represent three experiments. c Real-time PCR analysis of IL-8 was performed to determine remaining mRNA levels. Total RNA was prepared from BEAS-2B cells pre-stimulated with dTCTP (10 μg/ml) for 6 h followed by treatment with or without SB203580 (10 μM) for 30 min, and incubated with actinomycin D (1 μg/ml) for 0, 1, 2, 4 or 6 h. IL-8 mRNA levels of dTCTP-treated group without actinomycin D (0 h) were set as 100% IL-8 mRNA and relative percentages were calculated with respect to the 100% dTCTP control at other time points. Values are mean ± SEM (n = 3). *p < 0.05, **p < 0.01; significantly different from dTCTP treated group

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