Skip to main content
. 2014 Apr 15;158(3-4):224–232. doi: 10.1016/j.vetimm.2014.02.002

Fig. 2.

Fig. 2

Transient transfection reagents induce a type I interferon (IFN)-response in bovine monocyte-derived macrophages (bMDM) in the absence of siRNA. The siRNA uptake efficacy and off-target effects of five transfection reagents, INTERFERin (INT), Lipofectamine RNAiMAX (RNA), Lipofectamine 2000 (L2K), X-tremeGENE (XTG) and DharmaFECT 3 (DF3) were investigated. (A) bMDM were cultured with pre-determined concentrations of the transfection reagents in the absence of siRNA for 48 h (48 h incubation) or the medium was changed 24 h post transfection reagent treatment and cultured for a further 24 h (24 h incubation). The mRNA levels of the type I IFN-response gene interferon-induced protein with tetratricopeptide repeats 1 (IFIT1) were measured by qRT-PCR. The results are expressed as the IFIT1 mRNA fold difference compared to that measured in untreated cells (NC). Error bars denote the variation between three technical replicates. (B) bMDM were transfected with MEFV siRNA using the five transfection reagents for 24 h before the medium was replaced. MEFV mRNA levels in bMDM transfected with (NTC) and without (TC) non-target control siRNA using Lipofectamine RNAiMAX were also investigated. After 48 h the bMDM were activated with 100 ng/ml LPS for 2 h. The results are expressed as the percentage MEFV mRNA detected compared to that measured in NC. Error bars denote the variation between three technical replicates. The results are representative of three repeat experiments using bMDM from different animals.