Figure 3.
Effect of DHX9 shRNA on TLR-stimulated cytokine induction in human monocytes. A, THP-1 cells were transduced with lentiviruses encoding DHX9-targeting shRNA (A7 and G2) or a NSC shRNA. After transduction, the THP-1 cells were incubated in 1 μg/ml of puromycin for 1 week. Each THP-1 cell line was either left untreated (−) or incubated with 1 μg/ml of doxycycline (+) for 72 h to induce the required shRNA. Lysates were generated, resolved by SDS-PAGE, and immunoblotted for DHX9 and tubulin. B–E, doxycycline-treated THP-1 cells expressing shRNAs NSC, A7, or G2 were stimulated with 2 μg/ml of Pam3CSK4 (PAM3), 2.5 μg/ml of CLO75, 100 ng/ml of LPS or medium alone. The supernatants were removed 24 h later and TNFα (B), IL-6 (C), IP10 (D), and RANTES (E) secretions were measured by ELISA. The data are mean ± S.D. of triplicate samples and is representative of three experiments. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001 compared with NSC. N.D., not detected.