Induction of upregulated expression of TLR3 (A), TLR7 (B) mRNAs and TLR3 (C), TLR7 (D) proteins in P815 cells by GM-CSF. For real-time PCR analysis, cells were treated with various concentrations of GM-CSF at 37°C for 2, 6 and 16 h, respectively. The data were normalized to the housekeeping gene (β-actin gene) and were expressed as Mean ± SE fold over control for four separate experiments performed in duplicate. For flow cytometry analysis, the cells were incubated with various concentrations of GM-CSF for 16 h (C) or with 100 ng/ml of GM-CSF for 2, 6 and 16 h (D). ht-GM-CSF represented the heat-treated GM-CSF (100 ng/ml). Values shown are Mean ± SE for four separate experiments performed in duplicate. * P < 0.05 compared with the response to corresponding medium alone control.