Journal: BMC Complementary and Alternative Medicine

Article Title: Resveratrol distinctively modulates the inflammatory profiles of immune and endothelial cells

doi: 10.1186/s12906-017-1823-z

Figure Lengend Snippet: Effect of RES on gene expression in activated THP-1. PMA-treated THP-1 cells were cultured in the presence of indicated concentrations of RES and activated with LPS for 4 h. RT-PCR was performed and the gene expression levels were indicated as mean fold changes (± errors) (versus unstimulated cell) (see also reference ). * p < 0.05, ** p < 0.01, *** p < 0.001 (versus LPS-stimulated cells)

Article Snippet: THP-1 cells (from Cell Lines Service Eppelheim, Germany) were maintained at <2 × 10 5 cells/mL in RPMI 1640 medium supplemented with 50 U/mL penicillin, 50 μg/mL streptomycin, 10% FCS and 2 mM L-glutamine.

Techniques: Gene Expression, Cell Culture, Reverse Transcription Polymerase Chain Reaction

Journal: BMC Complementary and Alternative Medicine

Article Title: Resveratrol distinctively modulates the inflammatory profiles of immune and endothelial cells

doi: 10.1186/s12906-017-1823-z

Figure Lengend Snippet: Resveratrol reduced the secretion of cytokines and chemokines in LPS-activated THP-1 cells

Article Snippet: THP-1 cells (from Cell Lines Service Eppelheim, Germany) were maintained at <2 × 10 5 cells/mL in RPMI 1640 medium supplemented with 50 U/mL penicillin, 50 μg/mL streptomycin, 10% FCS and 2 mM L-glutamine.

Techniques:

Journal: Arthritis & Rheumatology (Hoboken, N.j.)

Article Title: Identification of Histone Deacetylase Inhibitor Targeting Type I Interferon and B Cell Abnormalities in Systemic Lupus Erythematosus

doi: 10.1002/art.43434

Figure Lengend Snippet: Experimental workflow and identification of vorinostat as an IFN‐I inhibitor. (A) Schematic overview. (B) Compounds were evaluated to reduce IFN‐I induction activity without affecting cell viability (reporter assay and cell counting kit‐8). (C) Top 20 compounds from screening showed potent IFN‐I inhibition with maintained viability. (D) Dose‐dependent IFN‐I inhibition by vorinostat. THP1‐Blue ISG cells were stimulated with LPS (500 ng/mL), 2'3'‐cGAMP (5 μg/mL), and R848 (5 μg/mL). ISG‐inducing activity (SEAP assay) decreased with vorinostat. (E) Validation using in PBMCs (same stimuli). IFN‐I activity (SEAP assay) decreased with vorinostat. Data represent the mean ± SEM from duplicate measurements across three independent experiments. IFN, interferon; LPS, lipopolysaccharide; NZB/W F1, New Zealand Black/White F1; PBMC, peripheral blood mononuclear cell; SAVI, STING‐associated vasculopathy with onset in infancy; SEAP, secreted embryonic alkaline phosphatase; SLE, systemic lupus erythematosus.

Article Snippet: THP1‐Blue ISG cells purchased from InvivoGen (San Diego, CA) and peripheral blood mononuclear cells (PBMCs) were seeded and treated with compounds from the Prestwick Chemical Library (Prestwick Chemical, San Diego, CA) at 1 μM in the presence of resiquimod (R848; 10 μg/mL), 2'3'‐cyclic GMP‐AMP (2'3'‐cGAMP; 5 μg/mL), or IFN‐I (Universal IFN‐I; 200 U/mL) for 24 hours.

Techniques: Activity Assay, Reporter Assay, Cell Counting, Inhibition, SEAP Assay, Biomarker Discovery

Journal: Nucleic Acids Research

Article Title: Insights into innate immune activation via PS-ASO–protein–TLR9 interactions

doi: 10.1093/nar/gkac618

Figure Lengend Snippet: Non-CpG PS-ASOs generate immune responses via the TLR9 pathway. ( A ) Model PS-ASOs. ‘o’ corresponds to phosphodiester linkage; all other linkages are phosphorothioate. Blue and orange indicate cEt and MOE 2′ modifications respectively. ( B ) Relative qRT-PCR levels of CCL22 mRNA in WT or TLR9-KO Bjab cells following 8 h incubation of indicated PS-ASOs at 1.6 μM in serum-free RPMI media by free-uptake. (right) Western blot analysis of TLR9 from indicated cells. ( C ) Relative fluorescence units (RFU) representing alkaline phosphatase activity under the NFKB promotor of HEK293 cells expressing hTLR9 (293-TLR9) showing TLR9 activation from 10 μM of the indicated PS-ASOs for 16 h. ( D ) RFU of TLR9 activation for 16 h with the indicated PS-ASOs in the following descending concentrations in μM: 10, 5, 2.5, 1.25, 0.62, 0.31, 0.156. Error bars are standard deviations from at least three independent experiments. P-values were calculated based on unpaired t-test and were computed comparing with control (WT or Luciferase) samples. *** P < 0.001; ** P < 0.01; * P < 0.05; ns, not significant.

Article Snippet: HEK-Blue™ hTLR9 cells (Invivogen) (293-TLR9) and THP1-Dual™ hTLR9 cells (Invivogen) (THP1-TLR9) are stable commercial cell lines that co-expresses the TLR9 gene and an optimized secreted embryonic alkaline phosphatase (SEAP) reporter gene in HEK293 or THP1 cells.

Techniques: Quantitative RT-PCR, Incubation, Western Blot, Fluorescence, Activity Assay, Expressing, Activation Assay, Luciferase