Journal: Microorganisms

Article Title: Monosodium Glutamate Inhibits Pseudomonas aeruginosa -Induced Acute Lung Injury by Targeting the Type III Secretion Systems and Modulating Host Immunity

doi: 10.3390/microorganisms14030725

Figure Lengend Snippet: KEGG pathway enrichment analysis and key protein expression in P. aeruginosa -induced acute lung injury (ALI) in mice. ( A ) KEGG pathway enrichment of differentially expressed genes between the ALI model and control groups; ( B ) protein levels of phospho-JNK, p-p38, p-p65, TLR4, and MyD88 in lung tissues compared between the ALI model and control groups by Western blot analysis (β-actin loading control); ( C ) comparison of protein levels (p-JNK, p-p38, p-p65, TLR4, MyD88) in lung tissues between the model and MSG-treated groups, as detected by Western blot (β-actin control) (n = 12 samples per group; data represent means ± SEM; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: The antibodies for phospho-NF-κB p65 (82335-1-RR), p-65 (80979-1-RR), phospho-P38 (14064-1-AP), phospho-JNK (80024-1-RR), MyD88 (23230-1-AP), TLR-4 (19811-1-AP), P38 (14064-1-AP), JNK (51153-1-AP), and β-actin (66009-1-Ig) were purchased from ProteinTech Group (Wuhan, China).

Techniques: Expressing, Control, Western Blot, Comparison

Journal: Journal of Biological Chemistry

Article Title: X-linked Inhibitor of Apoptosis Protein Functions as a Cofactor in Transforming Growth Factor-β Signaling

doi: 10.1074/jbc.m100331200

Figure Lengend Snippet: FIG. 1. XIAP associates with the TGF-b RI receptor. The indi- cated expression vectors were transiently transfected into 293 cells and coprecipitations were performed with glutathione-Sepharose beads, fol- lowed by immunoblotting with anti-HA antibody. The expression of the GST fusion proteins was also confirmed by using an anti-GST antibody (data not shown). A, coprecipitation of XIAP with the constitutively active mutants (denoted by *) of the TGF-b receptor superfamily. B, coprecipitation of XIAP with wild type, constitutively active, or kinase- deficient versions of TbRI.

Article Snippet: Cells were then incubated for 30 min at room temperature with either of the following primary antibodies: anti-hILP/XIAP mouse monoclonal antibody (Transduction Laboratories, San Diego, CA) or rabbit anti-HA polyclonal antibody (1:200, sc-805, Santa Cruz, Santa Cruz, CA).

Techniques: Expressing, Transfection, Western Blot

Journal: Journal of Biological Chemistry

Article Title: X-linked Inhibitor of Apoptosis Protein Functions as a Cofactor in Transforming Growth Factor-β Signaling

doi: 10.1074/jbc.m100331200

Figure Lengend Snippet: FIG. 2. XIAP colocalizes with TbRI. HepG2 cells were transfected with both XIAP and HA-TbRI expression vectors. Cells were immuno- stained with either anti-hILP/XIAP (mouse) or anti-HA (rabbit) anti- bodies followed by secondary staining with either fluorescein isothio- cyanate- or rhodamine-conjugated secondary antibodies as described under “Experimental Procedures,” and were visualized by fluorescence microscopy for XIAP and/or TbRI localization. The subcellular localiza- tion of XIAP is shown in green (top left), TbRI in red (top right), and areas of co-localization in yellow (bottom right). A representative field is shown.

Article Snippet: Cells were then incubated for 30 min at room temperature with either of the following primary antibodies: anti-hILP/XIAP mouse monoclonal antibody (Transduction Laboratories, San Diego, CA) or rabbit anti-HA polyclonal antibody (1:200, sc-805, Santa Cruz, Santa Cruz, CA).

Techniques: Transfection, Expressing, Staining, Fluorescence, Microscopy

Journal: Journal of Biological Chemistry

Article Title: X-linked Inhibitor of Apoptosis Protein Functions as a Cofactor in Transforming Growth Factor-β Signaling

doi: 10.1074/jbc.m100331200

Figure Lengend Snippet: FIG. 6. XIAP activation of JNK is inhibited by both DN Smad4 and DN TAK1. The indicated expression vectors were cotransfected with HA-JNK into 293 cells, and Jun kinase activity of JNK immunoprecipitates was assayed using a GST-c-Jun-(1–79) substrate (top panel) as described under “Experimental Procedures.” Lysates were immunoblotted with an anti-HA antibody (middle panel) to confirm equal loading. Complexes were evaluated by phosphorimage analysis (lower panel). XIAP was coexpressed with DN Smad4 (A) or DN TAK1 (B), or was treated with 50 mM SB203580 or 30 mM SB202190 for 24 h after transfection to block p38 (C), or alternatively the indicated XIAP deletion mutants were tested (D). All data are representative of at least three independent experiments.

Article Snippet: Cells were then incubated for 30 min at room temperature with either of the following primary antibodies: anti-hILP/XIAP mouse monoclonal antibody (Transduction Laboratories, San Diego, CA) or rabbit anti-HA polyclonal antibody (1:200, sc-805, Santa Cruz, Santa Cruz, CA).

Techniques: Activation Assay, Expressing, Activity Assay, Transfection, Blocking Assay

Journal: Journal of Biological Chemistry

Article Title: X-linked Inhibitor of Apoptosis Protein Functions as a Cofactor in Transforming Growth Factor-β Signaling

doi: 10.1074/jbc.m100331200

Figure Lengend Snippet: FIG. 8. XIAP does not co-precipitate with Smad4. The indicated expression vectors were transiently transfected into 293 cells, and co- precipitations were performed with glutathione-Sepharose beads fol- lowed by immunoblotting with a mixture of anti-Myc and anti-HA antibodies. The expression of the GST fusion proteins was also con- firmed by using an anti-GST antibody (data not shown).

Article Snippet: Cells were then incubated for 30 min at room temperature with either of the following primary antibodies: anti-hILP/XIAP mouse monoclonal antibody (Transduction Laboratories, San Diego, CA) or rabbit anti-HA polyclonal antibody (1:200, sc-805, Santa Cruz, Santa Cruz, CA).

Techniques: Expressing, Transfection, Western Blot