Figure 3. KLF5 mediates the pro-proliferative effect of oncogenic KRAS in IEC-6 cells.
IEC-iKRAS cells were induced with IPTG for 24 h and then transfected or not with KLF5 siRNA or non-specific (NS) siRNA. RNA and protein were extracted from cells 24 h after siRNA transfection. (A) Real-time PCR was performed on cDNA generated by KLF5-specific primers. The relative KLF5 mRNA levels were determined after adjusting for the levels of control β-actin, with the level in IPTG-treated iKRAS cells set at 1.0. Shown are the means of three independent experiments. (B) Proteins were analyzed by Western blotting using KLF5, KRAS and β-actin antibodies. The treatment conditions were identical to (A). (C) Cell proliferation was measured by cell counting. N = 6 and * p < 0.05, comparing between KLF5 siRNA3- and NS siRNA-treated cells. (D) Anchorage-independent colony formation in soft agar 21 days post-transfection. N = 6; * p < 0.01.