Figure 3.

Quantification of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and interleukin (IL)-10 mRNA. Quantitative, competitive reverse transcription–polymerase chain reaction (RT–qcPCR) was performed with (a) non-stimulated and (b) concanavalin A (Con A)-stimulated (24 hr) peripheral blood mononuclear cells (PBMC) from a naive pig. For PCR, cDNA and a 10-fold dilution of MIMIC were added together in concentrations ranging from 10¹ to 10⁻² amol for GAPDH (lanes 1–4, upper bands), and from 10² to 10⁻¹ amol for IL-10 (lanes 5–8, lower bands). In both Con A- and non-stimulated PBMC, molar equivalence was obtained in the presence of ≈ 10⁰−10¹ amol IL-10-specific MIMIC (lane 7, both panels) and 10⁻¹ amol GAPDH-specific MIMIC (lane 3, both panels). M, molecular-weight markers.