I have found in the literature that H154 is a specific inhibitor of TLR9. For this reason, I performed an experiment using 10uM H154 in macrophages (PMA-differentiated THP1 cells) before stimulation with 5uM CpG ODN 2216 (specific agonist of TLR9). In Figure 1 you can see the four conditions that I tested and the timepoints when I collected supernatant (3, 12, 24h).

I measured IFN beta using ELISA because this cytokine is suposed to be released after activation of TLR9 as one of the Type 1 IFNs.

The problem is that I could not see a real inhibition with H154 and I wondered which could be the problem (Figure 2).

Do you have any suggestions?

Thank you in advance!

H154 sequence: C*C*T*C*A*A*G*C*T*T*G*A*G*G*G*G

*Phosphorothioate bond.

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