Figure 4

Inhibition of transcriptional targets of 5’-DFUR and gemcitabine by AQP3-siRNA. Non-transfected (white), negative control siRNA (dashed) or AQP3-siRNA (black) transfected MCF7 cells were incubated for 90 minutes with 250 μM 5’-DFUR or 100 nM gemcitabine. After 24 h, RNA was isolated and real-time RT-PCR analysis for p21 and Fas performed (a and b, respectively). C_T values for each gene are normalized to an endogenous reference gene (GAPDH). mRNA levels are calculated in arbitrary units using control values as the reference. Results are presented as means ± SE of four to six independent experiments measured in duplicate. Statistical significance was assessed with the Student’s t test (* p < 0.05; ** p < 0.01). (c) Non-transfected or AQP3-siRNA transfected MCF7 cells were treated for 90 minutes with 250 μM 5’-DFUR or 100 nM gemcitabine and Western blot analysis was perfomed for p21, Fas and α-tubulin as a loading control. A representative Western blot is shown.