Human TRIF/TICAM1 Antibody

Detection of Human TRIF/TICAM1 by Western Blot. Western blot shows lysates of Ramos human Burkitt's lymphoma cell line and Raji human Burkitt's lymphoma cell line. PVDF Membrane was probed with 1 µg/mL of Mouse Anti-Human TRIF/TICAM1 Monoclonal Antibody (Catalog # MAB6216) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for TRIF/TICAM1 at approximately 113 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

Detection of Human TRIF/TICAM1 by Western Blot MyD88 is required for inflammasome sensing of HIV and HCV.Knockdowns of the TLR adaptors MyD88 (A, B) and TICAM-1 (TRIF, E, F) were generated and confirmed by previously described RNA interference techniques. (*) denotes comparisons with p≤0.05 compared to the mock transfected cells. Monocytes in which MyD88 was knocked down were cultured with HIVBaL (solid bars) or HCVSubject 180 (hatched bars) and pro-IL-1 beta mRNA transcription measured at 6 h (C, G) and IL-18 secretion measured at 24 h (D, H). Shown are the relative production of pro-IL-1 beta mRNA and IL-18 in MyD88 (C, D) or TICAM-1 (G, H) knockdown monocytes normalized to mock transfected monocytes (no siRNA) stimulated with the same viruses. Bars represent the mean ± S.D. for n = 6–9 independent transfection experiments, (**) denotes comparisons with p≤0.001 compared to the scramble siRNA transfected cells. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.ppat.1004082), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human TRIF/TICAM1 by Western Blot Differential importance of endosomal TLRs in inflammasome sensing of HIV and HCV.Monocytes were cultured without stimulation or with HIVBaL or HCVSubject 180 and IL-1 beta, IFN alpha and measured at multiple timepoints. Fold change in expression of IL-1 beta (black bar) and IFN alpha (grey bar) following HIVBaL or HCVSubject 180 relative to stimulation with media alone is shown at 6 h (A). Functional knockdowns of the endosomal located TLRs in monocytes were generated by RNA interference. Monocytes were mock transfected or transfected with either siRNA targeting TLR7 or a non-targeting sequence (Scramble). After 24 h, cell lysates were prepared and efficacy of knockdown determined by (B) western blot and (C) qRT-PCR. (D) Specificity of the TLR7 siRNA was confirmed by qRT-PCR using primers for TLR3, TLR7, TLR8, and TLR9. (*) denotes comparisons with p≤0.05 compared to the mock transfected cells. Monocytes in which endosomal TLR knockdown were generated were cultured with HIVBaL (solid bars) or HCVSubject 180 (hatched bars) and pro-IL-1 beta mRNA transcription measured at 6 h (E–H) and IL-18 secretion measured at 24 h (I–L). Shown are the relative production of pro-IL-1 beta mRNA and IL-18 in TLR8 (E, I), TLR7 (F, J), TLR3 (G, K) and TLR9 (H, L) knockdown monocytes normalized to mock transfected monocytes (no siRNA) stimulated with the same viruses. Bars represent the mean ± S.D. for n = 6–9 independent transfection experiments, (**) denotes comparisons with p≤0.05 and (***) denoted p≤0.001 compared to the scramble siRNA transfected cells. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.ppat.1004082), licensed under a CC-BY license. Not internally tested by R&D Systems.