Type I Interferons Function as Autocrine and Paracrine Factors to Induce Autotaxin in Response to TLR Activation
Fig 5
The synergistic effect of IFN-γ on the ATX induction by IFN-β.
(A) THP-1 cells were treated for 4h with IFN-β (10 ng/ml) and/or IFN-γ (50 ng/ml) as indicated. ATX mRNA levels were detected by qRT-PCR. (B) THP-1 cells were stimulated by LPS for 16 h with or without IFN-γ priming. ATX mRNA levels were detected by qRT-PCR. (C) THP-1 cells were preincubated with IFN-β specific neutralizing antibody (anti-IFN-β; 1μg/ml) or negative control antibody (rabbit IgG; 1 μg/ml) for 30 min, and then subjected to LPS and/or IFN-γ treatment as indicated. ATX mRNA levels were detected by qRT-PCR after treatment for 16 h. (D) IFNAR1 siRNA and non-specific siRNA (siNC) were transfected into THP-1 cells respectively. After siRNA transfection for 48 h, THP-1 cells were treated with LPS plus IFN-γ for 16 h. IFNAR1 were detected by Western blot, and ATX mRNA expression was analyzed by qRT-PCR. The ATX expression detected by qRT-PCR analyses was normalized to expression of GAPDH and presented relative to expression in untreated cells. All qRT-PCR data are expressed as mean values ± SD, n = 3. The p values derived from Student’s t test are (*) p < 0.05, (**) p < 0.01. A representative experiment out of three is shown.