Figure 3
Effects of different pathways on cell number, Mmp13 and Tnfa expression. (a) MTT analysis was performed by spectrophotometry of chondrocytes cultured with vehicle (control) or 10 ng/ml TGFα in the presence or absence (vehicle) of 10 μM ROCK (Y27632), MEK (U0126), p38 MAPK (SB202190) or PI3K (LY294002) inhibitor. The mean absorbance readings (plus s.d.) from N=5 independent cell isolations are shown. Significant reductions compared with vehicle only occurred in all inhibitor-treated samples +/−TGFα (statistics not shown for simplicity). (b) Real-time PCR was performed using RNA samples isolated from chondrocytes incubated with vehicle (control) or 10 ng/ml TGFα for 48 h in the presence or absence of 10 μM inhibitors as described above. Mean gene expression values (plus s.d.) for matrix metalloproteinase-13 (Mmp13) and (c) tumor necrosis factor-α (Tnfa) from N=4 independent cell isolations are shown normalized to controls. Significantly different (P<0.05) TGFα-treated vs control means are indicated by asterisks.
