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. 2022 Jan 21;12:658895. doi: 10.3389/fimmu.2021.658895

Figure 2.

Figure 2

(A) Wt BMDMs were stimulated with 0.5 µM ssRNA40-chol or ssRNA40 complex to DOTAP or 0.5 µM dsRNA40-chol, dsRNA40 complexed to DOTAP and dsRNA40-chol-2’OM. Medium, CpG2216 (1 µM), LPS (1 µg/ml) and DOTAP served as controls. Supernatants were taken after 20 h of incubation and cytokine production was analyzed by ELISA. Data were calculated from three individual experiments. Bars indicate mean + SEM. Unpaired t-test was used for comparison of ssRNA40 samples while one-way-ANOVA with Sidak’s post-hoc test was used for comparison of dsRNA40 samples. *p < 0.05, **p < 0.01, ****p < 0.0001. (B) Purified human Monocytes were isolated via MACS separation, incubated with (blue bars) or without (black bars) 2 µM of the TLR8 inhibitor Cu-CPT9a and stimulated with dsRNA40-chol, dsRNA40 complexed to DOTAP and dsRNA40-chol-2’OM at 2.5 µM, 0.5 µM and 0.1 µM. Medium, CpG2216 (1 µM), LPS (1 µg/ml), ssRNA/DOTAP(2,5µM), R848 (2,5 µg/ml), TL8-506 (60 nM) and DOTAP served as controls. Supernatants were harvested after 20 h of incubation and IL-6, TNF-α, IFN-α, IFN-β were analyzed by ELISA. Data were calculated from at least three (for TL8-506) to six independent experiments. Bars indicate mean + SEM. *p < 0.05, ***p < 0.001, ****p < 0.0001; two-way ANOVA with Sidak’s post-hoc test.