tlr agonist r848  (InvivoGen)

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Reprogramming tumor microenvironment via systemic delivery of TLR3 agonist and manganese nanoparticle

doi: 10.1073/pnas.2409559121

Figure Lengend Snippet: Mn 2+ augments IFN-I responses of TLR agonists. ( A ) Mouse BMDCs were incubated with various concentrations of Mn 2+ with or without 10 µg/mL of each TLR agonist, and after 24 h, IFN-β in the media was quantified. ( B ) CT26 tumor-bearing mice were treated by i.t. administration with 10 µg of each TLR agonist with or without 2.5 µg Mn 2+ on days 10, 13, 16, and 19, followed by monitoring of tumor growth. ( C ) Proinflammatory cytokine release from mouse BMDCs treated with either LMW-poly(I:C) (10 µg/mL), Mn 2+ (0 to 500 µM), or their combination. ( D ) Mouse BMDCs were incubated with either 5 µg/mL LMW-poly(I:C), 500 µM Mn 2+ , or their combination for 8 h, followed by immunoblotting for marker proteins in the STING-IFN-I pathway. ( E ) Relative densitometric values of Western blots as measured by ImageJ (NIH). The data represent mean ± SEM from a representative experiment of two independent experiments; n = 4 ( A and C ) and n = 3 to 5 ( B ) biologically independent samples. Two independent experiments were analyzed for ( E ). The data were analyzed by two-way ANOVA ( B and C ) or one-way ANOVA ( E ) with Bonferroni’s multiple comparisons test.

Article Snippet: Mouse BMDCs (1 × 10 5 /well) were seeded in a 96-well plate and incubated with various concentrations of Mn 2+ (Sigma Aldrich, no. 244589) with or without 10 μg/mL of each TLR agonist (Invivogen, no. tlrl-picw, tlrl-pic, tlrl-mdp, tlrl-pms, vac-mpla2, tlrl-r848-1 or Integrated DNA Technologies, no. 01181197Q).

Techniques: Incubation, Western Blot, Marker

Journal: PLOS Pathogens

Article Title: Peculiar transcriptional reprogramming with functional impairment of dendritic cells upon exposure to transformed HTLV-1-infected cells

doi: 10.1371/journal.ppat.1012555

Figure Lengend Snippet: a. Schematic representation of the experimental design. MDDCs were exposed to uninfected (Jurkat, green) or HTLV-1-infected T cells (C91-PL, blue) for 24h, before restimulation with LPS or R848 for an additional 24h. b. Flow cytometry analysis after CD11c and CD86 staining, in LPS- ( left ) or R848-restimulated MDDCs ( right ) pre-exposed to Jurkat (red) or C91-PL (dark blue) cells. Data are represented as the normalized MFI of CD86, with the MFI in restimulated Jurkat-pre-exposed MDDCs set to 100 (n = 30 or 10 independent experiments, respectively). Presented data are a subset of , respectively and were analysed with Kruskal-Wallis test or ordinary one-way ANOVA, respectively as described in and Tables c. Supernatant from the indicated cocultures was collected after LPS restimulation, and TNF-α ( left ) and IFN-I ( right ) concentrations were quantified for n = 3 or 10 independent experiments, respectively. Presented data are a subset of and were analysed with RM one-way ANOVA or ordinary one-way ANOVA, respectively as described in and Tables. d. Schematic drawing summarizing the results from Figs and . The drawing was created using BioRender.com .

Article Snippet: Toll-like receptor (TLR)-4 agonist (LPS, tlrl-3pelps; 1μg/mL) and TLR-7/8 agonist (R848, tlrl-r848; 3μg/mL) were purchased from Invivogen.

Techniques: Infection, Flow Cytometry, Staining