Induction of MMP-1 protein by tenascin-C is ERK1/2, JNK and p38 dependent.

<p>(a) ASM cells were stimulated with soluble tenascin-C (10 µg/ml). Lysates were harvested at 20 minutes and assayed by immunoblot for a panel of phosphorylated signalling proteins using proteome profiler. (b) ASM cells from three normal donors were stimulated with soluble tenascin-C (10 µg/ml). Lysates were harvested at 20 minutes and 24 hours, with levels of total and phosphorylated signalling proteins assayed by Western Blotting. (c) ASM cells from three normal donors were incubated for 30 minutes with inhibitors of JNK (SP600125, 10 µmol), MEK1/2 (U0126, 10 µmol), Rac1 (NS23766, 100 µmol), p38 (SB208350, 10 µmol), mammalian target of rapamycin complex 1 (rapamycin 10 nmol), PI3K (wortmanin 100 nmol) or glycogen synthase kinase αβ (GSK3αβ, SB216763 10 µmol) prior to stimulation with tenascin-C (10 µg/ml). Supernatant was harvested after 24 hours and assayed for MMP-1 protein. Expression levels were normalised to vehicle control (0.02% DMSO, tenascin-C (10 µg/ml)) and analysed using a 1-way ANOVA with <i>post-hoc</i> Dunnett’s multiple comparison test (*P≤0.05. ***P<0.001).