Skip to main content
. 2014 Jan 3;9(1):e84513. doi: 10.1371/journal.pone.0084513

Figure 2. IL2/PHA PBMCs fail to induce type I IFN responses and pro-inflammatory cytokines upon DNA transfection.

Figure 2

(A–D) IL2/PHA PBMCs were transfected with HIV-derived ssDNA and dsDNA (2 µg/mL), or infected with SeV (MOI 0.5). Total RNA was isolated after 6 hours for RT-qPCR measurements of (A) IFNβ, (B) CXCL10, (C) ISG56, and (D) Viperin. (E–H) IL2/PHA PBMCs were transfected with ssDNA and dsDNA (2 µg/mL), or infected with SeV (MOI 0.5). Supernatants were harvested after 24 hours and analyzed for CXCL10, TNFα, MIP-1α, and IL6 protein levels. (I–J) PMA (100 nM) treated THP-1 cells were transfected with ssDNA and dsDNA (2 µg/mL). Total RNA was harvested after 6 hours for RT-qPCR measurements of IFNβ and CXCL10 mRNA. (K) Primary human monocyte-derived macrophages (MDM)s were transfected with ssDNA and dsDNA (2 µg/mL). Total RNA was harvested after 6 hours for RT-qPCR measurements of IFNβ. (L) Un-stimulated PBMCs were isolated and immediately transfected with ssDNA and dsDNA (2 µg/mL), or infected with SeV (MOI 0.5). Supernatants were harvested after 24 hours and analyzed for CXCL10 by ELISA. Both PCR and ELISA data are shown as means of triplicates +/− SD. Similar results were obtained in three independent experiments. Mock, Lipofectamine.