Figure 1

Mycoplasma synoviae and its diacylated lipopeptide induce expression of TLR15 in chicken macrophages. A) Expression of TLR15, TLR1 and TLR2 mRNA after infection of HD11 cells with Mycoplasma synoviae. HD11 cells were infected with live Mycoplasma synoviae (MS) for 1 h, 6 h or 24 h and mRNA expression was analyzed by RT-qPCR. NC represents non-infected cells. Bars show mean ± S.E. (n = 4); ***, p < 0.001 compared to non-infected cells. B) Expression of TLR15, TLR1 and TLR2 mRNA after treatment of HD11 cells with diacylated lipopeptide or its non-acylated analog. HD11 cells were treated with 1 μM non-acylated peptide (NAP) or diacylated lipopeptide (MDLP) for 1 h, 6 h or 24 h and mRNA expression was analyzed by RT-qPCR. NC represents non-treated cells. Bars show mean ± S.E. (n = 4); **, p < 0.01 and ***, p < 0.001 compared to NAP-treated cells at each time point for selected gene. C) Expression of TLR15, TLR1 and TLR2 mRNA after treatment of MDM cells with diacylated lipopeptide or its non-acylated analog. MDM cells were treated with 1 μM non-acylated peptide (NAP) or diacylated lipopeptide (MDLP) for 1 h, 6 h or 24 h and mRNA expression was analyzed by RT-qPCR. NC represents non-treated cells. Bars show mean ± S.E. (n = 3); *, p < 0.05 and **, p < 0.01 compared to NAP-treated cells at each time point for selected gene. D) Expression levels of TLR15 and β-actin, as determined by Western blotting. HD11 cells were treated with NAP or MDLP for 24 h and TLR15 protein levels were determined. Levels of endogenous β-actin were determined by Western blotting to validate input for each sample.