Figure 4

Validation of the two-dimensional-PAGE data by quantitative RT-PCR and Validation of the protein pathway analysis data. A, expression of selected genes in hVSMCs treated with TNF-α or melittin for 12 h was determined by quantitative RT-PCR. Total RNA was isolated from hVSMCs, reverse transcribed and amplified with the specific primers indicated in under “materials and methods.” β-actin was used as the control. B, Total cell lysates (25 μg) and nuclear lysates of hVSMCs treated for 12 h with either TNF-α or melittin were separated by SDS-PAGE. Proteins were blotted onto a PVDF membrane, probed with specific antibodies, and detected as described under “materials and methods.”