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. 2009 Sep 30;284(47):32305–32311. doi: 10.1074/jbc.M109.008250

FIGURE 2.

FIGURE 2.

AMF induces IL-8 protein production and IL-8 mRNA transcription. A, mean IL-8 protein concentrations ± S.E. (n = 3) detected by ELISA in cell culture media in response to stimulation with rAMF (500 ng/ml). SBcl-2 and 1205Lu cells were incubated with various concentrations of rAMF for 12 h. B, mean IL-8 protein concentrations ± S.E. (n = 3) detected by ELISA in cell culture media from untreated SBcl-2 cells and SBcl-2 cells stimulated with rAMF (500 ng/ml). SBcl-2 cells were incubated with or without rAMF and extracted at various times until 96 h. C, representative RT-PCR amplification of IL-8. SBcl-2 cells were stimulated with rAMF (500 ng/ml) and collected after 1, 2, 4, 6, and 12 h. C, control samples. D (top), IL-8 mRNA stability. SBcl-2 cells were exposed to rAMF for 4 h and then maintained in fresh medium containing 5 μg/ml ActD (−AMF +ActD) or 5 μg/ml ActD plus 500 ng/ml rAMF (+AMF +ActD). The cells were harvested at different time points. Total RNA was extracted, and the IL-8 and β-actin (control) mRNA levels were detected by RT-PCR. D (bottom), mean IL-8 mRNA ratios (n = 3) measured by real-time RT-PCR in cells undergoing the protocols described for the top. The mRNA levels were expressed as percentages relative to those measured after 4 h of rAMF treatment.