Figure 4

IL-1β and iFGFR1 induce expression of Cox-2 in HC-11/R1 cells. (a) HC-11/R1 cells were treated with 5 ng/ml rmIL-1β for the indicated times. Quantitative RT-PCR was used to analyse IL-1β mRNA expression (normalised to levels of cyclophilin and glyceraldehyde 3-phosphate dehydrogenase (GAPDH)). Error bars represent standard error of the mean. ** P < 0.01. (b) HC-11/R1 cells were treated with either 30 nM AP20187 or ethanol (Sol) for the indicated times and cyclooxygenase (Cox) 2 mRNA expression was analysed and normalised as described in a. * P < 0.05, ** P < 0.01. (c) HC-11/R1 cells were stimulated with either 5 ng/ml recombinant murine (rm) IL-1β or 30 nM AP20187 for the indicated times. Immunoblot analysis was performed using antibodies specific for Cox-2 and β-tubulin as a loading control. Densitometry was performed to analyse Cox-2 expression levels relative to β-tubulin. (d) HC-11/R1 cells were treated with 30 nM AP20187 and 5 ng/ml rmIL-1β for one hour. Quantitative RT-PCR analysis was performed to examine levels of Cox-2 expression, which were normalised to levels of cyclophilin and GAPDH. Error bars represent standard error of the mean. ** P < 0.01, *** P < 0.001. (e) Migration assays were performed as described in Figure 3. Cells were treated with 30 nM AP20187 and/or 5 ng/ml IL-1β in the presence of either celecoxib or dimethyl sulfoxide (DMSO) as a solvent control as indicated. Error bars represent standard error of the mean. * P < 0.05, ** P < 0.01. iFGFR1 = inducible fibroblast growth factor receptor 1.