Fig. 3.

Confirmation of selected genes by qRT-PCR. Macrophages (3 × 10⁶/ml) were incubated with live B. burgdorferi (Bb) spirochetes at an MOI of 10, sonicated (son) spirochetes (1 × 10⁷/ml), or L-OspA (1 μg/ml) in the presence (black bars) or absence (stimulant; white bars) of 10 ng/ml of mouse recombinant IL-10 (rIL-10). Controls consisted of cells incubated with rIL-10 or medium alone (unstimulated cells). RNA samples were collected after 24 h of incubation, and gene transcripts were quantified by TaqMan qRT-PCR. All values were normalized with respect to the housekeeping gene Gapdh mRNA levels. Results are presented as fold increase over control (the level in unstimulated cells). Asterisks indicate significant differences from cells incubated with stimulants alone (P < 0.05). P values were calculated by an unpaired Student's t test. Results are representative of one of two experiments. Each bar represents the mean ± SD from samples run in duplicates.