Figure 3

ILK regulates CCL2 expression. A. Si-RNA was used to knockdown ILK and a scrambled oligonucleotide was used as a control. RNA was extracted from HCT116 cells after 4 h of stimulation with IL-1β and reverse transcribed to make cDNA. This was used to determine the message of the molecules shown using RT-PCR. CCL2 induction in response to the cytokine IL-1β was the only molecule whose expression was blocked by si-RNA knockdown of ILK (see CCL2 panel, lane 4). This result was reproduced on 2 further occasions. B. The experiment was repeated as in A except Q-PCR used to determine the expression of CCL2, either following si-RNA knockdown of ILK (left panel) or using the QLT0267 inhibitor (right panel). C. HCT116 cells were pretreated with the specific ILK inhibitor QLT0267 for 1 hour before being stimulated with IL-1β for the times indicated. CCL2 concentrations were determined in the culture supernatent using ELISA. The data are representative of two separate experiments, each performed in triplicate (*p < 0.01).