Journal: Molecular Cell

Article Title: Human NLRP1 is a sensor of pathogenic coronavirus 3CL proteases in lung epithelial cells

doi: 10.1016/j.molcel.2022.04.033

Figure Lengend Snippet:

Article Snippet: Nigericin , tlrl-nig-5 , Invivogen.

Techniques: Virus, Variant Assay, Clinical Proteomics, Recombinant, Microscopy, Plasmid Preparation, Mutagenesis, Construct, Software

Journal: Neuroscience

Article Title: Protection of granulocyte-colony stimulating factor to hemorrhagic brain injuries and its involved mechanisms: effects of vascular endothelial growth factor and aquaporin-4.

doi: 10.1016/j.neuroscience.2013.12.017

Figure Lengend Snippet: Fig. 9. Effects of signal pathway inhibitors on VEGF and AQP4 expression. (A) ELISA showed that VEGF protein was up-regulated by rhG-CSF, which was suppressed by STAT3 inhibitor AG490 (n = 6, ⁄p < 0.05 vs. control, #p < 0.05 vs. G-CSF). (B–I) Both Western blot and immunofluorescence revealed that rhG-CSF increased AQP4 protein expression in astrocytes, which was inhibited by U0126, an ERK inhibitor (n = 6, ⁄p < 0.05 vs. control, #p < 0.05 vs. G-CSF). (B) Western blot analysis. (C) Group of control. (D) Group of G-CSF. (E) Group of G-CSF plus U0126, an ERK inhibitor. (F) Group of G-CSF plus SB239063, a p38-MAPK inhibitor. (G) Group of G-CSF plus SP600125, a JNK inhibitor. (H) Group of G-CSF plus AG490, a STAT3 inhibitor. (I) Group of G-CSF plus Ly294002, a PI3K inhibitor. Nuclei are highlighted by DAPI staining. Scale bar = 50 lm.

Article Snippet: Group 3: G-CSF plus U0126 (10 lmol/L, Invivogen, San Diego, CA, USA), an ERK inhibitor.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Control, Western Blot, Staining

Journal: iScience

Article Title: Heat-killed Mycobacterium tuberculosis induces trained immunity in vitro and in vivo administered systemically or intranasally

doi: 10.1016/j.isci.2024.108869

Figure Lengend Snippet:

Article Snippet: CpG ODN 2216 , InvivoGen , Cat# tlrl-2216.

Techniques: Blood/Plasma/Serum Samples, Recombinant, Saline, Enzyme-linked Immunosorbent Assay, Isolation, Reverse Transcription, SYBR Green Assay, Lactate Assay, Staining, Software

Journal: Cell Research

Article Title: The RNA-binding protein Mex3B is a coreceptor of Toll-like receptor 3 in innate antiviral response

doi: 10.1038/cr.2016.16

Figure Lengend Snippet: Mex3B regulates poly(I:C)-triggered signaling through TLR3. (A) Mex3B(G83/177D) inhibits poly(I:C)- but not TRIF-induced signaling. The 293 cells (1 × 10 5 ) were transfected with the indicated plasmids and increased amounts of a Mex3B plasmid. Twenty-four hours after transfection, cells were treated with poly(I:C) (20 μg/ml) for 6 h before luciferase assays were performed. (B) TLR3(C95/122A) inhibits poly(I:C)-induced, TLR3/Mex3B-mediated signaling. The experiments were similarly performed as in A . (C) Knockdown of Mex3B inhibits poly(I:C)- but not TRIF- or TBK1-mediated activation of the IFN-β promoter. The 293-TLR3 cells (1 × 10 5 ) were transfected with the IFN-β promoter reporter (0.1 μg) and either a control or Mex3B-RNAi plasmid (0.5 μg). Transfected cells were selected with puromycin (1 μg/ml) for 24 h followed by either treatment with poly(I:C) (20 μg/ml) or retransfection with TRIF or TBK-1 plasmids (0.1 μg). Luciferase assays were performed 6 h after treatment or 18 h after retransfection. Data are represented as mean ± SEM. * P < 0.05, ** P < 0.01 (Student's t -test). (D) Effects of chloroquine on TLR3- and Mex3B-mediated signaling. The 293 cells (1 × 10 5 ) were transfected with the indicated plasmids (0.1 μg each) for 24 h. The cells were pretreated with chloroquine for 30 min, and then treated with poly(I:C) (20 μg /ml) for 6 h before luciferase assays.

Article Snippet: Poly(I:C), LPS, R837, PGN, chloroquine (Invivogen); TNFα, IL-1β (R&D Systems); D-galactosamine (Sigma); GM-CSF (peproTech); ELISA kit for murine IFN-β, TNFα, IL-6 (Biolegend); dual-specific luciferase assay kit (Promega); EZ-link Psoralen-PEG 3 -Biotin (Thermo); streptavidin agarose (Solulink); lysosome isolation kit (sigma); mouse monoclonal antibodies against Flag, β-actin (Sigma), HA (OriGene), phospho-IBα (Cell Signaling Technology), TLR3 (IMGENEX); rabbit monoclonal antibodies against TLR3, phospho-IRF3 (Cell Signaling Technology); rabbit polyclonal antibodies against IRF3 (Santa Cruz Biotechnology), TRIF (Cell Signaling Technology), TBK1 and phospho-TBK1(EPITOMICS), Flag (MBL); Alexa Fluor 555-conjugated goat anti-rabbit IgG antibody, Alexa Fluor 647-conjugated goat anti-mouse IgG antibody (Invitrogen) were purchased from the indicated manufacturers.

Techniques: Transfection, Plasmid Preparation, Luciferase, Knockdown, Activation Assay, Control