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. 2015 Mar 10;10(3):e0119727. doi: 10.1371/journal.pone.0119727

Fig 5. Macrophage-like differentiated THP-1 cells respond to endosomal TLR ligands but not to the canonical TLR13 ligand.

Fig 5

(A, B) THP-1 cells were either treated with 10 nM PMA or left untreated (as described in Material and Methods) before stimulation with R848 (5 μg/ml), LTA (5 μg/ml), LPS (10 ng/ml) or DOTAP-mediated delivery of poly(I:C) (5 μg). Supernatants were collected after 8 h and IL-8 (A) or TNF (B) release was determined by ELISA. Error bars represent SDs (n≥3). (C, D) RNA was isolated from differentiated or undifferentiated THP-1 cells treated as described in (A) and (B) for 4 or 8 h. IL8 (C) and TNF (D) mRNA levels were determined by qRT-PCR (normalized to HPRT). Mean values ± SD are shown (n ≥ 3). (E) THP-1 cells stably expressing TLR13 (TLR13 THP-1) were stimulated with TLR13 ligand SA19, poor TLR13 ligand (mut-SA19), S. pyogenes RNA or LTA for 4 h. TNF was determined in supernatants by ELISA. Error bars in represent SDs (n≥3).