Figure 9

MEK and PI3K inhibition blocks MØCM induced growth, but dissociates kinase activity from cyclin D1 expression. A-B: (A) LM2 or (B) JF32 cells were cultured alone (Control), with 2 ng/mL mIGF-1 (+ IGF-1) or with media conditioned by MH-S macrophages (+ MØCM), and treated with 0.05% DMSO (vehicle, "Veh"), 5 μM U0126 (U0), 10 μM LY294002 (LY) or both agents together (U + L). Relative cell number was determined by MTS, and mean + SD plotted as fold-change from vehicle control, which was normalized to 1.0. Data was pooled from at least 3 independent experiments. * P < 0.05, ** P < 0.01 and *** P < 0.001 versus control for each treatment by 2-way ANOVA. C-D: LM2 (C) or JF32 (D) cells were cultured as described above, and cell homogenates probed for protein expression as described. Western blot images are representative of two independent experiments. E-I: densitometry data from all western blot replicates was collected from both LM2 and JF32 cell lines, combined, and presented as fold change (mean ± SEM) from untreated vehicle control lanes at each time point ("Veh -" group normalized to 1, not shown). The legend colors indicate drug treatment groups, followed by a "- or +" to indicate the absence or presence of MØCM, as in (A-B). * P < 0.05, ** P < 0.01 and *** P < 0.001 versus control (Veh -) cells at each time point by 2-way ANOVA.