Journal: Biomedicines
Article Title: TIRAP/Mal Positively Regulates TLR8-Mediated Signaling via IRF5 in Human Cells
doi: 10.3390/biomedicines10071476
Figure Lengend Snippet: TIRAP silencing attenuates cytokine production from MDMs challenged with clinical isolates of E. coli (ECO), while affecting mainly IL-12A induction by Group B streptococcus (GBS) and pro-inflammatory cytokine induction by S. aureus (SAU). MDMs (5–6 donors in consecutive experiments) were pre-treated with TIRAP siRNA or control oligo and incubated with LPS (100 ng/mL), CL075 (1 µg/mL), or live bacteria (GBS 248, SAU 17-2, and ECO 18-1) for a total time of four hours. The doses of bacteria were 1 × 10 5 /mL (e5) and 1 × 10 6 /mL (e6). This roughly corresponds to MOI 0.01 and 0.1 for GBS, MOI 0.02 and 0.2 for SAU, and MOI 0.1 and 1.0 for ECO. Gene expression was determined by RT-qPCR, normalized to untreated sample, and presented as a mean relative fold change +SEM. Statistical testing was done with 2-way RM-ANOVA and post-test (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).
Article Snippet: Ultrapure K12 LPS from E. coli , thiazoloquinoline compound CL075, and synthetic diacylated lipoprotein FSL-1 (Pam2CGDPKHPKSF) were from InvivoGen (San Diego, CA, USA).
Techniques: Control, Incubation, Bacteria, Gene Expression, Quantitative RT-PCR