FIGURE 2: TL-532 induces the best biological balance between inflammation in RAW264.7 wt cells and tumor inflammatory cell death in NCI-H292 wt cells. (A, D) Schematic representation of the 70 bp dsRNAs showing their differing (A:U) and (I:C) bp content. (B, E) Activation of myeloid cells by TLR3-ligands was determined on RAW264.7 cells treated with the different molecules at 10 µg/mL for 24 hours. The concentration of mTNFα was measured by ELISA. (C, F, G, H) Activation of tumor cells by TLR3-ligands was determined on NCI-H292 cells treated with the different molecules at 500 µg/ml (C), 100 µg/ml (F, G), or 10 µg/ml (H) for 24 hours. Cell viability was measured with MTS assay and expressed as percentage of untreated cells (C, G). Apoptosis was measured with AnnexinV-Glo assay and expressed as fold increase of untreated cells (F). The concentration of hIL-6 was measured by ELISA (H). (B, C) Among the tested molecules, TL-532 and TL-533 induce the highest biological responses in both RAW264.7 wt and NCI-H292 wt cells. (E) Increasing (I:C) bp content reduces RAW264.7 wt cells activation. TL-536 is inactive in RAW264.7 wt, unlike TL-532. (F-H) Increasing (I:C) bp content does not impact the induction of apoptosis, viability reduction, and hIL-6 secretion in NCI-H292 wt cells except for TL-540, which induces less hIL-6 secretion comparatively. TL-536 is barely active in NCI-H292 wt cells, contrary to the TL-532. Data are the representative (C) or are the mean of three independent experiments (B, E-H). Results are expressed as mean ± SD. Unpaired Student's t-test: * = p<0.05; ** = p<0.01; *** = p<0.001; ns: not significant. Unpaired Student's t-test values are compared to Mock condition unless otherwise stated.