Journal: Molecular medicine reports
Article Title: Expression of glycoprotein nonmetastatic melanoma protein B in macrophages infiltrating injured mucosa is associated with the severity of experimental colitis in mice.
doi: 10.3892/mmr.2015.4408
Figure Lengend Snippet: Figure 4. Reduced Gpnmb expression increases cytokine and chemokine production and activates p38 and ERK1/2 signaling. (A) RAW264.7 murine macro phage cells were transfected with NC‑siRNA, Gp‑siRNA, or vehicle alone, and the expression of Gpnmb was examined using RT‑qPCR. Data are presented as the mean ± SD (n=3; *P<0.001). (B) Effects of Gpnmb knockdown on levels of mRNAs encoding IL‑1β, IL‑6, TNF‑α and MCP‑1 were examined using RT‑PCR. Data are presented as the mean ± SD (n=3; *P<0.01; **P<0.001. (C) Effects of Gpnmb knockdown on p38, ERK1/2, and JNK signaling and IκB activation were examined using western blotting. (D) Expression of Gpnmb in the RAW264.7 cells was decreased in response to LPS and increased following the removal of LPS. RAW264.7 macrophage cells were exposed to LPS (100 µg/ml) for 24 h and changes in Gpnmb were examined using RT‑qPCR. mRNA expression levels in the cells without LPS for 6 h were set as 1. Data are presented as the mean ± SD (n=3). *P<0.05; **P<0.01; ***P<0.001. Gpnmb, glycoprotein nonmetastatic melanoma protein B; IL, interleukin; TNF, tumor necrosis factor; MCP, monocyte chemoattractant protein; LPS, lipopolysaccharide; ERK, extracellular signal‑regulated kinase; NC, negative control; Gp, Gpnmb; siRNA, small interfering RNA; RT‑qPCR, reverse transcription‑quantitative poly merase chain reaction; JNK, c‑Jun N‑terminal kinase; p‑, phosphorylated.
Article Snippet: Polyclonal rabbit anti-mouse phosphorylated (p)-ERK1/2 (1:1,000, cat. no. 9101), ERK1/2 (1:1,000, cat. no. 3372), phosphorylated c-jun N-terminal kinase (p-JNK; 1:1,000; cat. no. 9251), p-p mitogen activated kinase (MAPK; 1:1,000; cat. no. 4511), p38 MAPL (1:1,000, cat. no. 9212) and IκB (1:1,000; cat. no. 9242), as well as monoclonal rabbit anti-mouse JNK (1:1,000; cat. no. 9258) were purchased from Cell Signaling Technology Inc. (Danvers, MA, USA).
Techniques: Expressing, Transfection, Knockdown, Activation Assay, Western Blot, Negative Control, Small Interfering RNA
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