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. 2004 Jun;24(11):4743–4756. doi: 10.1128/MCB.24.11.4743-4756.2004

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FIG. 7. — Dex induces MKP-1. (A) MKP-1 mRNA was measured by real-time quantitative reverse transcription-PCR in A549 cells after 8 h of treatment with the indicated concentrations of Dex. Asterisks denote a significant increase between untreated and Dex-treated cultures as determined by Tukey-Kramer pair comparison analysis (P < 0.05). (B) Kinetics of MKP-1 induction by Dex. The MKP-1 mRNA level was measured by real-time quantitative reverse transcription-PCR. A549 cells were treated with 100 nM Dex and harvested at the indicated time points (0, 1, 2, 4, 6, 8, and 24 h). Asterisks denote a significant increase between untreated and Dex-treated cultures as determined by Tukey-Kramer pair comparison analysis (P < 0.05). (C) The GR antagonist, RU486, counteracts the Dex-induced up-regulation of the MKP-1 mRNA level. A549 cells were stimulated with Dex for 8 h, and the MKP-1 mRNA levels were measured by real-time quantitative reverse transcription-PCR. RU486 counteracts the enhancing effect of Dex on MKP-1 mRNA levels. For each well, MKP-1 expression was normalized to cyclophilin B, and the fold induction for each experiment was determined by dividing the normalized expression from treated wells by that from control (untreated) wells. The asterisk denotes a statistically significant increase between the indicated Dex-treated wells and all other treatments as determined by Tukey-Kramer pair comparison analysis (P < 0.05). Values are the mean ± SE of three experiments. (D) MKP-1 protein levels in A549 cells after Dex treatment. MKP-1 protein expression was detected after Dex treatment. Western blot (IB) analysis was performed to confirm the Dex-induced effect. A549 cells were harvested after 8 h of treatment and immunoprecipitated (IP) with anti-MKP-1 antibody. The graph shows MKP-1 protein expression from cells treated with Dex and/or RU486, normalized to the control immunoprecipitated protein (vehicle treated cells). Analysis of the immunoreactive bands with NIH-Image software reproduced the Dex-induced increase in the level of MKP-1. Values are the mean ± SE of three experiments. *, P < 0.05 for pair comparison analysis (Tukey-Kramer test).

FIG. 7. — Dex induces MKP-1. (A) MKP-1 mRNA was measured by real-time quantitative reverse transcription-PCR in A549 cells after 8 h of treatment with the indicated concentrations of Dex. Asterisks denote a significant increase between untreated and Dex-treated cultures as determined by Tukey-Kramer pair comparison analysis (P < 0.05). (B) Kinetics of MKP-1 induction by Dex. The MKP-1 mRNA level was measured by real-time quantitative reverse transcription-PCR. A549 cells were treated with 100 nM Dex and harvested at the indicated time points (0, 1, 2, 4, 6, 8, and 24 h). Asterisks denote a significant increase between untreated and Dex-treated cultures as determined by Tukey-Kramer pair comparison analysis (P < 0.05). (C) The GR antagonist, RU486, counteracts the Dex-induced up-regulation of the MKP-1 mRNA level. A549 cells were stimulated with Dex for 8 h, and the MKP-1 mRNA levels were measured by real-time quantitative reverse transcription-PCR. RU486 counteracts the enhancing effect of Dex on MKP-1 mRNA levels. For each well, MKP-1 expression was normalized to cyclophilin B, and the fold induction for each experiment was determined by dividing the normalized expression from treated wells by that from control (untreated) wells. The asterisk denotes a statistically significant increase between the indicated Dex-treated wells and all other treatments as determined by Tukey-Kramer pair comparison analysis (P < 0.05). Values are the mean ± SE of three experiments. (D) MKP-1 protein levels in A549 cells after Dex treatment. MKP-1 protein expression was detected after Dex treatment. Western blot (IB) analysis was performed to confirm the Dex-induced effect. A549 cells were harvested after 8 h of treatment and immunoprecipitated (IP) with anti-MKP-1 antibody. The graph shows MKP-1 protein expression from cells treated with Dex and/or RU486, normalized to the control immunoprecipitated protein (vehicle treated cells). Analysis of the immunoreactive bands with NIH-Image software reproduced the Dex-induced increase in the level of MKP-1. Values are the mean ± SE of three experiments. *, P < 0.05 for pair comparison analysis (Tukey-Kramer test).