Figure 3. Suppression of PIASy expression down-regulates Ets-1 expression.

COS-7 cells were transiently transfected with either control siRNA or PIASy siRNA. At 48 h after transfection, cells were treated with vehicle (DMSO) as a control or MG132 (20 μM). After 5 h, cell lysates were prepared and subjected to immunoblotting with PIASy-, Ets-1- and β-actin-specific antibodies (top panel). Immunoblot band densities were determined using ImageJ (NIH), and PIASy and Ets-1 levels were normalized to the loading control, β-actin (middle and bottom panels respectively). The value for the control siRNA transfected cells in the absence of MG132 was assigned an arbitrary value of 1.0.