Figure 6

Involvement of RELA and RELB in gene activation in response to extracellular and intracellular polyIC treatments. HT1080 cell line was treated with 40 μg/ml of ex-polyIC and 0.4 μg/ml of in-polyIC for 8 hours after knocking down RELA and RELB using gene-specific siRNAs. Gene expression was measured using qPCR. Graphs show the average of two independent experiments, and the student's t-test was performed to indicate statistically significant differences between non-specific siRNA control and gene-specific knock-down cells. (* is for P-value≤0.05, ** is for P-value≤0.01 and NS is 'not significant'.) A. Nucleofection of RELA and RELB siRNAs achieved about 92% and 60% knock-down efficiencies, respectively. B. IFNB, IL8 and CCL3 inductions in response to in-polyIC treatments are dependent on RELA but independent on RELB. C. IFNB, TNF and CCL3 inductions in response to ex-polyIC treatment are independent of RELA but dependent on RELB. Particularly, IFNB, TNF and CCL3 inductions in response to ex-polyIC treatment are significantly increased in RELB knock-down. D. HT1080 cell line with RELB-knocked-down was treated with 40 μg/ml of ex-polyIC in the absence/presence of 10 μg/ml cycloheximide. The increase in gene inductions of IFNB, TNF and CCL3 is diminished by cycloheximide treatment.