Ligands for TLR-4 and TLR-9 induce prolonged pro-inflammatory gene transcription in draining LNs. RNA was isolated from the LN whole-cell population cryopreserved with TRIzol reagent on days 1 and 3 after TLR-L injection, and were analyzed by low-density array quantitative real-time PCR for a panel of 93 genes involved in immune responses. All genes and time points were first normalized to the average cycling threshold value of expression of the housekeeping genes for 18s ribosomal RNA, Actb (β-actin), and Gusb (β-glucuronidase). Transcripts were grouped by their immune function and origin and their expression was evaluated at the time points matching the innate cell kinetics in blood. (A) Heat map of gene transcripts. Each row represents a mean –fold change relative to baseline at the indicated time point after TLR-L injection in each of the experimental groups. The 23 genes with a significant change from baseline at any given time point and experimental group are shown. Kinetics of expression of the ISGs Isg-15, Ifit-1, Mx-1, and Oas-1 (B); the selected chemokines: Cxcl-10 (IP-10), Cxcl-11 (I-TAC), Ccl-3 (MIP-1α), and Ccl-2 (MCP-1) (C); and complement genes: C3 and C3ar1 (D) are represented as the relative change of mRNA copies at the indicated time points after TLR-L injection compared with baseline. Dotted line marks the cutoff (y = 1) of the fold expression change. Data are means ± SEM. *P < .05; **P < .01; ***P < .001 by ANOVA.