Journal: Cell reports

Article Title: Candida albicans oscillating UME6 expression during intestinal colonization primes systemic Th17 protective immunity

doi: 10.1016/j.celrep.2022.110837

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Anti-mouse TLR4/MD2 (clone MTS510) , Hycult Biotech , Cat#HM1029; RRID:AB_533218.

Techniques: Recombinant, Staining, Conjugation Assay, SYBR Green Assay, Software

Journal:

Article Title: Lipopolysaccharide-Trap-Fc, a Multifunctional Agent To Battle Gram-Negative Bacteria

doi: 10.1128/IAI.00004-09

Figure Lengend Snippet: LPS-Trap-Fc1 to -Fc4 bind LPS and block LPS-mediated IL-6 production in Mono Mac 6 cells. (A) Supernatants of HEK293T cells transfected with LPS-Trap-Fc1 were incubated in the presence (lane 1) or absence (lane 3) of biotin-LPS. Additionally, supernatants were preincubated with an anti-TLR4/MD-2 MAb (lane 2) or an excess of LPS (lane 4). Complexes were immunoprecipitated with streptavidin-Sepharose, subjected to SDS-PAGE, and analyzed by Western blotting with an anti-FLAG MAb. (B) RAW 264.7 cells were incubated with LPS-Trap-Fc1 or a medium control. Cells were stimulated with 100 ng/ml LPS overnight, and IL-6 levels in supernatants were determined by ELISA. *, P < 0.002 compared to control (Mann-Whitney U test, n = 6).

Article Snippet: For specificity control, some supernatants were preincubated with either 50 μg/ml LPS ( Salmonella enterica serovar Minnesota; Sigma), which has a high binding activity for TLR4/MD-2, or 10 μg/ml anti-mouse TLR4/MD-2 MTS510 MAb (Hycult Biotechnology, Uden, The Netherlands) for 1 h at room temperature.

Techniques: Blocking Assay, Transfection, Incubation, Immunoprecipitation, SDS Page, Western Blot, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal:

Article Title: Hyperplastic gastric tumors induced by activated macrophages in COX-2/mPGES-1 transgenic mice

doi: 10.1038/sj.emboj.7600170

Figure Lengend Snippet: Stimulations of TLR on the gastric epithelial cells with bacterial PGN or LPS. (A) RT–PCR for TLR2 and TLR4 mRNAs using total RNA from gastric mucosa sampled by LMD (top) and primary culture of the epithelial cells (bottom), respectively. COX-2 in LMD-sample RT–PCR was used as positive control. GAPDH was used as endogenous controls. WT, wild type; TG, K19-C2mE. Gastric epithelial cells were treated with PGN (B) and LPS (C) for 20 h. Concentrations of TNF-α in the supernatants are presented as the mean±s.d. RAW264 mouse monocyte cells were used for positive control. For TLR4 blocking, anti-TLR4 antibody was added at 10 μg/ml (blue bars). *P<0.05.

Article Snippet: For blocking the TLR4 signaling, monoclonal antibody to mouse TLR4/MD2 (Clone MTS510; HyCult biotechnology, Uden, Netherlands) was used at 10 μg/ml.

Techniques: Reverse Transcription Polymerase Chain Reaction, Positive Control, Blocking Assay