Figure 2

Effects on S100A4 expression and MTX sensitivity upon siS100A4 transfection of HT29 cells. A) HT29 cells (30,000) were transfected with siS100A4 as described in Methods. Total RNA was extracted after 48 h and S100A4 mRNA levels were determined by RT-Real-Time PCR. B) S100A4 protein levels were determined by Western Blotting 72 h after transfection, using specific antibodies against S100A4 and Actin to normalize the results. C) Chemosensitization assays toward methotrexate: cells were treated with siS100A4 for 48 h and then incubated with MTX. Cell viability was determined 3 days after MTX treatment. The expression and viability results are expressed as percentages referred to the untreated cells. Values are the mean of three independent experiments ± SE. A representative image of Western Blots is presented. *p < 0.05, ** p < 0.01, *** p < 0.001.