Journal: Biomolecules

Article Title: Comparison of In Vitro Multiple Physiological Activities of Cys–Tyr–Gly–Ser–Arg (CYGSR) Linear and Cyclic Peptides and Analysis Based on Molecular Docking

doi: 10.3390/biom16010126

Figure Lengend Snippet: Cell viability of RAW 264.7 macrophages treated with L-CR5 and C-CR5.

Article Snippet: The mouse-derived macrophage cell line RAW 264.7 and the human dermal fibroblast cell line CCD-986sk were obtained from the Korean Cell Line Bank (KCLB, Seoul, Republic of Korea).

Techniques:

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Adiponectin and heme oxygenase-1 suppress TLR4/MyD88-independent signaling in rat Kupffer cells and in mice after chronic ethanol exposure

doi: 10.4049/jimmunol.1002060

Figure Lengend Snippet: A and B) Kupffer cells isolated from pair- and ethanol-fed rats were cultured with 1μg/ml globular adiponectin (gAcrp) for 18h. Cells were then stimulated with 100 ng/ml LPS for 4h. The expression of IFN-β and CXCL10 mRNA was normalized to 18S mRNA. Values are expressed relative to Kupffer cells from pair-fed rats not treated with gAcrp. n = 4, * p < 0.05 ethanol-fed compared with pair-fed, +p < 0.05 compared with LPS-stimulated cells not treated with gAcrp. C-F) RAW 264.7 cells were cultured for 18h in the absence or presence of increasing doses of either gAcrp (0–1 μg/ml) or full-length adiponectin (flAcrp, 0–10 μg/ml). Cells were then stimulated with 100 ng/ml LPS for 4h and expression of IFN-β and CXCL10 mRNA was normalized to 18S mRNA. n = 4, + p < 0.05 compared with LPS-stimulated cells not treated with adiponectin. G and H) RAW 264.7 cells were cultured for 18h in the absence or presence of 10 μ/ml gAcrp or full-length adiponectin. Cells were then stimulated with 100 ng/ml LPS for 8h and the concentration of CXCL10 protein accumulated in the media was measured by ELISA. n = 6, + p < 0.05 compared with LPS-stimulated cells not treated with adiponectin.

Article Snippet: Mouse RAW 264.7 cell nucleofection kit was purchased from Lonza (Cologne, Germany).

Techniques: Isolation, Cell Culture, Expressing, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Adiponectin and heme oxygenase-1 suppress TLR4/MyD88-independent signaling in rat Kupffer cells and in mice after chronic ethanol exposure

doi: 10.4049/jimmunol.1002060

Figure Lengend Snippet: RAW 264.7 cells were transfected or not with 100 nM of AdipoR1 siRNA, AdipoR2 siRNA or scrambled siRNA and then cultured with or without 1 μg/ml gAcrp or 10 μg/ml flAcrp for 18h. Cells were then stimulated with LPS for 4h. IFN-β and CXCL10 mRNA was normalized to 18S mRNA. n=4, Values with different superscripts are significantly different from each other, p < 0.05.

Article Snippet: Mouse RAW 264.7 cell nucleofection kit was purchased from Lonza (Cologne, Germany).

Techniques: Transfection, Cell Culture

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Adiponectin and heme oxygenase-1 suppress TLR4/MyD88-independent signaling in rat Kupffer cells and in mice after chronic ethanol exposure

doi: 10.4049/jimmunol.1002060

Figure Lengend Snippet: A and B) Kupffer cells from pair- and ethanol-fed rats were isolated and cultured overnight with 0–1000 ng/ml gAcrp for 18 h. Expression of TLR4 and CD14 mRNA was normalized to 18S mRNA. Values are expressed as fold increases relative to Kupffer cells from pair-fed rats not treated with gAcrp. n = 4, * p < 0.05 ethanol-fed compared to pair-fed, +p < 0.05 compared with cells not treated with gAcrp. C) TLR4 promoter activity was measured in RAW 264.7 macrophages transiently transfected with a full-length TLR4 promoter-luciferase reporter construct and a Renilla luciferase control plasmid. After 24 h, cells were treated with or without 1 μg/ml gAcrp for 18 h. Values represent relative luciferase activity. n = 4; + p < 0.05. D) RAW 264.7 macrophages were treated without or with 1 μg/ml gAcrp for 18h and expression of TLR4 mRNA measured and normalized to 18S mRNA. n=4, + p <.05 compared with cells not treated with adiponectin. E) RAW 264.7 macrophages were cultured with or without 1 μg/ml gAcrp for 18h and cell surface expression of TLR4-MD2 (solid line), relative to isotype controls (dotted line), was measured by flow cytometry. Data are representative of four experiments.

Article Snippet: Mouse RAW 264.7 cell nucleofection kit was purchased from Lonza (Cologne, Germany).

Techniques: Isolation, Cell Culture, Expressing, Activity Assay, Transfection, Luciferase, Construct, Plasmid Preparation, Flow Cytometry

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Adiponectin and heme oxygenase-1 suppress TLR4/MyD88-independent signaling in rat Kupffer cells and in mice after chronic ethanol exposure

doi: 10.4049/jimmunol.1002060

Figure Lengend Snippet: A and B) RAW 264.7 macrophages were treated with or without 1 μg/ml gAcrp for 18h. HO-1 mRNA expression was normalized to 18S mRNA. B) HO-1 protein expression was measured by Western blot and normalized to total ERK1/2. n=4, * p < 0.05 compared with cells not treated with adiponectin C-F) RAW 264.7 macrophages were transfected with an empty vector or an HO-1 overexpression plasmid. Twenty four hour after transfection, cells were treated or not with 1 μg/ml gAcrp for 18h. C) TLR4 mRNA was measured and normalized to 18S mRNA. n=4, * p < 0.05 compared to non-treated cells. D and E) Transfected cells were then stimulated with LPS for 4h. IFN-β (D) and CXCL10 (E) mRNA were measured normalized to 18S mRNA. n=4, * p < 0.05 compared with LPS-stimulated cells not treated with gAcrp or HO-1 overexpression. F) Transfected cells were then stimulated with LPS for 2h and Western blot analysis was performed using antibodies to phosphorylated STAT1 and Hsc70 (loading control). Images are representative of four experiments.

Article Snippet: Mouse RAW 264.7 cell nucleofection kit was purchased from Lonza (Cologne, Germany).

Techniques: Expressing, Western Blot, Transfection, Plasmid Preparation, Over Expression

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Adiponectin and heme oxygenase-1 suppress TLR4/MyD88-independent signaling in rat Kupffer cells and in mice after chronic ethanol exposure

doi: 10.4049/jimmunol.1002060

Figure Lengend Snippet: A) RAW 264.7 cells were transfected or not with 100 nM of HO-1 siRNA or scrambled siRNA and then cultured with or without 1 μg/ml gAcrp for 18h. Cells were then stimulated with LPS for 4h. IFN-β and CXCL10 mRNA was normalized to 18S mRNA. n=4, Values with different superscripts are significantly different from each other, p < 0.05. B) RAW264.7 cells were cultured with or without 0.5 μM ZnPP in the presence and absence of 1 μg/ml gAcrp for 18h. Cells were then stimulated with LPS for 4h. IFN-β and CXCL10 mRNA was normalized to 18S mRNA. n=4, Values with different superscripts are significantly different from each other, p < 0.05.

Article Snippet: Mouse RAW 264.7 cell nucleofection kit was purchased from Lonza (Cologne, Germany).

Techniques: Transfection, Cell Culture

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Adiponectin and heme oxygenase-1 suppress TLR4/MyD88-independent signaling in rat Kupffer cells and in mice after chronic ethanol exposure

doi: 10.4049/jimmunol.1002060

Figure Lengend Snippet: A and B) RAW 264.7 cells were cultured for 18h in the absence or presence of either gAcrp (1 μg/ml), CoPP (10μM) or CORM2 (100 μM). Cells were then stimulated without or with 100 ng/ml LPS for 4h and expression of IFN-β and CXCL10 mRNA normalized to 18S mRNA. n = 4, +p < 0.05 compared with LPS-stimulated cells not treated with inducers of HO-1. C and D) Kupffer cells isolated from pair- and ethanol-fed rats were cultured with 10μM CoPP for 18h. Cells were then stimulated with 100 ng/ml LPS for 4h. Expression of IFN-β and CXCL10 mRNA was normalized to 18S mRNA. Values are expressed relative to Kupffer cells from pair-fed rats not treated with gAcrp. n = 4, * p < 0.05 ethanol-fed compared with pair-fed, +p < 0.05 compared with LPS-stimulated cells not treated with CoPP.

Article Snippet: Mouse RAW 264.7 cell nucleofection kit was purchased from Lonza (Cologne, Germany).

Techniques: Cell Culture, Expressing, Isolation