FIGURE 5.

MS-275 and IGFBP-3 modulate p21 expression and cell migration. Cells were transfected with scrambled or IGFBP-3 siRNA and incubated for 24 h and then changed to fresh media containing vehicle or 6 μm MS-275 for an additional 24 h. p21 mRNA was analyzed by qRT-PCR (A) and p21 protein by Western blot (B) with actin as loading control. Data are shown as mean fold change ± S.D. (triplicates) relative to the scrambled siRNA-transfected cells without MS-275 treatment and are representative of three experiments. All qRT-PCR values were normalized against HMBS. *, p < 0.001 versus scrambled siRNA. C, migration of cells transfected with control or IGFBP-3 siRNA was analyzed as described under “Experimental Procedures.” Left panel, representative images of one of three fields per insert by phase contrast microscopy. Right panel, absorbance at 560 nm of dye extracted from stained migrated cells. Data are shown as means ± S.D. (triplicates) from one experiment representative of three. *, p < 0.005 versus scrambled siRNA (ANOVA).