Figure 7

Participation of the 2-5A system in the poly(I:C) transfection-induced antitumor effect. (a) One day after poly(I:C) transfection (500 ng/ml), total RNA was extracted, and 4 μg total RNA from each sample was denatured and analyzed by electrophoresis using 1% agarose-formaldehyde gels, ethidium bromide, and UV light. Similar results were obtained from two independent experiments. M, marker. (b) RCC cells were then transfected with either RNaseL or control siRNA. After 72 h, selective knockdown was confirmed by immunoblotting. α-tubulin was used as a control. (c) RCC cells, which were pre-transfected with the indicated siRNAs 72 h prior, were transfected additionally with poly(I:C) (SKRC-1, 250 ng/ml; SKRC-44, 31.3 ng/ml). Cell viability was determined using a WST-8 assay after 24 h. Similar results were obtained from three independent experiments. (d) RCC cells, which were pre-transfected with the indicated siRNA 72 h prior, were transfected additionally with poly(I:C) (SKRC-1, 500 ng/ml; SKRC-44, 50 ng/ml). The proportion of apoptotic cells was determined by flow cytometry after 24 h. Similar results were obtained from three independent experiments. pIC-TF, poly(I:C) transfection. *p < 0.05, **p < 0.01.