Figure 7.

Involvement of defined TLRs in the formation of TNTs derived from monocytes. PMHs were exposed to different TLR agonists (LPS-RS, Zymosan, CL097, and PAM, as indicated), and continuously monitored as described in the Figure 1. (A) 10 μg/mL LPS-RS, a TLR4 antagonist, was added to PHMs 15 min prior to exposure to 10 ng/mL LPS. A similar procedure was followed with 10 μg/mL LPS-RS and 3 μg Zymosan, an insoluble preparation of cell wall from Saccharomyces cerevisiae, recognized by TLR2 and also by Dectin-1. (B) 1 μg/mL of the synthetic bacterial lipopeptide PAM3CSK4 (PAM), a TLR2/1 ligand, and 3 μg/mL of the imidazoquinoline compound CL097, a TLR7/8 ligand, were applied. Images show TNT abundance at peak density time point (4–8 h). All images are representative of at least two independent experiments performed in duplicate. Scale bar is 50 μm.