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Figure 4. PKC-dependent COX-2 mRNA expression and MAPKs phosphorylation in odontogenic keratocyst fibroblasts. (A) Fibroblasts (1.5 x 10⁶ cells) were incubated without (Co) or with 0.1 nM rhIL-1 (IL-1) for 30 sec at 37°C. The amounts of Ins(1,4,5)P₃ in the cells (n = 4) were measured as described in "MATERIALS & METHODS". (B) Fibroblasts (4 x 10⁴ cells/cm²) were stimulated with 0.1 nM rhIL-1 for 10, 30, and 60 min. Then, the activity of PKC for the cells (n = 3) was measured as described in "MATERIALS & METHODS". Vertical bars indicate mean ± SD. * Significant difference at p < 0.05. (C) Fibroblasts (4 x 10⁴ cells/cm²) were stimulated with 0.1 nM rhIL-1 (lanes 2,3), 0.01% DMSO (lane 4), or 1 µM PMA (lane 5,6) for 6 hrs after pre-incubation without (lanes 1,2,4,5) or with (lanes 3,6) 2 µM staurosporine for 1 hr at 37°C. Then, the expression of COX-2 mRNA was measured as described in "MATERIALS & METHODS". (D) Fibroblasts (4 x 10⁴ cells/cm²) were pre-incubated in the absence (lanes 1,2) or presence (lane 3) of 2 µM staurosporine for 1 hr at 37°C. The cells were then stimulated with 0.1 nM rhIL-1 (lanes 2,3) for 10 min for measurement of the phosphorylation of ERK1/2 and p38, or for 20 min for measurement of the phosphorylation of JNK. The aliquots of cell lysates were subjected to Western immunoblotting as described in "MATERIALS & METHODS".

J DENT RES, Vol. 86, No. 2, 186-191 (2007)
DOI: 10.1177/154405910708600215

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