Figure 4

Effect of SMC3 on the expression of a set of early-response genes. a) Cells were transfected with 1 μg/ml of pcDNA3.1 (control) or SMC3-pcDNA3.1 and RNA collected after 48 hrs in 1 ml TriReagent. Gene transcripts were amplified by RT-PCR and the products analyzed by agarose electrophoresis. b) Analysis of RhoB level in NIH-3T3 and 293 cells overexpressing SMC3. Cells were transfected as in a) and 48 h later solubilized in lysis buffer. Twenty and 50 μg respectively of NIH-3T3 and 293 cell lysate proteins were analyzed by 12.5% SDS-PAGE. After transfer to nitrocellulose, RhoB was immunodetected using a rabbit polyclonal antibody followed by ECL detection of the immunocomplex. Nitrocellulose-bound proteins were stained with Ponceau-red to evidence the amount of proteins loaded.