Supplementary Figure 3: TIPARP-mediated regulation of type I IFN response to stimulation with viral mimetic nucleic acids.

(a) HEK293T cells expressing control plasmid (Control) or TIPARP were treated with 3pRNA for the indicated time periods, and then assayed for IFNB1 and IFNA1 mRNA expression by qRT-PCR. n = 3 per group. (b) Luciferase activity of a p-125Luc reporter plasmid after 24 h of 3pRNA stimulation in HEK293T cells transfected with control plasmid (C) or the increasing doses of TIPARP expression vector (0.01, 0.1, and 1 μg). n = 3 per group. (c) Quantitative RT-PCR analysis of IL8 mRNA expression levels after 24 h of IL-1β stimulation in HEK293T cells transfected with control (C) or the increasing doses (0.01, 0.1, and 1 μg) of TIPARP expression vector. n = 3 per group. (d) Knockdown efficiency of siTIPARP-1 was evaluated by qRT-PCR analysis of TIPARP mRNA expression levels in HEK293T cells (left). Quantitative RT-PCR analysis of IFNB1 mRNA induction by 3pRNA stimulation in HEK293T cells treated with siControl or siTIPARP-1 (right). n = 3 per group. (e) Quantitative RT-PCR analysis of Ifna1 mRNA induction in response to the indicated viral MAMPs in Tiparp+/+ or Tiparp−/− MEFs. n = 3 per group. (f) Tiparp+/+ or Tiparp−/− BMDMs were stimulated with poly(rI:rC) without any transfection reagent for 24 h and IFN-β production was measured by ELISA. In this case, poly(rI:rC) was used as a ligand for TLR3. n = 3 per group. **P < 0.01 as compared with control (Student’s t-test). Data are representative of at least two independent experiments with similar results (mean ± s.d.). ND, not detected. NS, not significant.