Journal: Marine Drugs

Article Title: Emerging Marine Biotoxins in European Waters: Potential Risks and Analytical Challenges

doi: 10.3390/md20030199

Figure Lengend Snippet: Recent in vitro methodology for emerging marine toxins identification.

Article Snippet: In parallel, a solid-phase receptor-based method for the detection of CI using a microsphere-flow cytometry system (Luminex) was developed.

Techniques: In Vitro, Binding Assay, Cytometry, Concentration Assay, Enzyme-linked Immunosorbent Assay, Immunodetection, Electrochemiluminescence, Cell Based Assay, Reporter Assay, Inhibition, Enzyme Immunoassay

Journal: Cell Cycle

Article Title: Inflammatory stimuli promote growth and invasion of pancreatic cancer cells through NF-κB pathway dependent repression of PP2Ac

doi: 10.1080/15384101.2015.1127468

Figure Lengend Snippet: Inflammatory stimuli promoted growth and invasion of pancreatic cancer cells in vitro. (A) MTT assay of viability of cells incubated with LPS (0, 10, 100, 1000 ng/ml) for 24, 48, and 72 h. (B) Cell invasion assay following 24-h exposure to 10 ng/ml LPS. Cells that had migrated to the lower membranes were photographed under ×400 magnification. **P < 0.01 as compared to control. (C) Quality control of MCM. After 7-day MCSF treatment, IL-8 and TNF-α levels at 24, 48, 72, 96, and 120 h were measured using Milliplex assay. (D) MTT assay of viability of cells incubated with MCM for 24, 48, 72, 96, and 120 h. *P < 0.05 as compared with control. (E) Cell invasion assay following 24-h exposure to MCM. Cells that had migrated to the lower membranes were photographed under ×400 magnification. **P < 0.01 as compared to control.

Article Snippet: Multiplex biometric immunoassay kits containing fluorescent dyed microbeads (Milliplex NF-κB Signaling Magnetic Bead kit; Cat. No. 48-630MAG, and Human Cytokine MAGNETIC Kit; Cat. No. HCYTOMAG-60K, Millipore Corp, St Charles, MO) were used for measuring phosphorylated IKKα/β (Ser176/Ser180), phosphorylated IκBα (Ser32), c-Myc, IL-8, and TNF-α.

Techniques: In Vitro, MTT Assay, Incubation, Invasion Assay, Control

Journal: Cell Cycle

Article Title: Inflammatory stimuli promote growth and invasion of pancreatic cancer cells through NF-κB pathway dependent repression of PP2Ac

doi: 10.1080/15384101.2015.1127468

Figure Lengend Snippet: Inflammatory stimuli induced NF-κB pathway activation in pancreatic cancer cells. (A, B) Milliplex assay evaluation of IKK phosphorylation at Ser176/Ser180 in SW1990 and CFPAC-1 cells treated with 10 ng/ml LPS (A) or MCM (B) for 12 h. (C) DN-IKKα (S176/180A) overexpression repressed LPS-induced IKK phosphorylation in SW1990 (top) and CFPAC-1 (bottom) cells. *P < 0.05, **P < 0.01 as compared with control. &P < 0.05 indicates significant differences between fold induction. (D) DN-IKKα (S176/180A) overexpression repressed MCM-induced IKK phosphorylation in SW1990 cells. *P < 0.05, **P < 0.01 as compared with control. &P < 0.05 indicates significant differences between fold induction. (E, F) Milliplex assay evaluation of IκBα phosphorylation at Ser32 after 12-h treatment with 10 ng/ml LPS (E) or MCM (F). (G) DN-IKKα (S176/180A) or DN-IκBα (S32/36A) overexpression repressed LPS-induced IκBα phosphorylation in SW1990 (top) and CFPAC-1 (bottom) cells. *P < 0.05, **P < 0.01 as compared with control. &P < 0.05, and&P < 0.01 indicate significant differences between fold induction. (H) DN-IKKα (S176/180A) or DN-IκBα (S32/36A) overexpression repressed MCM-induced IκBα phosphorylation in SW1990 cells. *P < 0.05, **P < 0.01 as compared with control. &P < 0.05, and&P < 0.01 indicate significant differences between fold induction. (I) Western blot of p65 expression in cells treated with 10 ng/ml LPS for 12 h, followed by total and nuclear protein extraction. β-actin and histone H1 were the internal controls for the total and nuclear extracts, respectively. (J, K) Luciferase reporter gene assays of cells transiently transfected with pNF-κB-luc before 36-h treatment with 10 ng/ml LPS (J) or MCM (K). **P < 0.01 as compared with control. (L, M) Milliplex assay of c-Myc expression in cells treated with LPS (L) or MCM (M) for 12 h. **P < 0.01 as compared with control.

Article Snippet: Multiplex biometric immunoassay kits containing fluorescent dyed microbeads (Milliplex NF-κB Signaling Magnetic Bead kit; Cat. No. 48-630MAG, and Human Cytokine MAGNETIC Kit; Cat. No. HCYTOMAG-60K, Millipore Corp, St Charles, MO) were used for measuring phosphorylated IKKα/β (Ser176/Ser180), phosphorylated IκBα (Ser32), c-Myc, IL-8, and TNF-α.

Techniques: Activation Assay, Phospho-proteomics, Over Expression, Control, Western Blot, Expressing, Protein Extraction, Luciferase, Transfection