Journal: BioMed Research International

Article Title: TNF α Mediated IL-6 Secretion Is Regulated by JAK/STAT Pathway but Not by MEK Phosphorylation and AKT Phosphorylation in U266 Multiple Myeloma Cells

doi: 10.1155/2013/580135

Figure Lengend Snippet: Regulation of IL-6 release by TNF α in vitro. After serum starvation, U266 and IM9 multiple myeloma cells were treated with or without 1 ng/mL TNF α for indicated times. Cell supernatants from each experimental sample were harvested for ELISA and IL-6 concentrations were determined at different time points with TNF α treatment. Relative fold changes compared to TNF α nontreated samples as a control were shown in graph. Bars represent the mean ± SEM from three independent experiments.

Article Snippet: IM9 was obtained from the Korean Cell Line Bank (Chongro-gu, Seoul, Republic of Korea).

Techniques: In Vitro, Enzyme-linked Immunosorbent Assay, Control

Journal: BioMed Research International

Article Title: TNF α Mediated IL-6 Secretion Is Regulated by JAK/STAT Pathway but Not by MEK Phosphorylation and AKT Phosphorylation in U266 Multiple Myeloma Cells

doi: 10.1155/2013/580135

Figure Lengend Snippet: Effect of TNF α on activation of MAPK, JNK, and PI3K/AKT signaling pathways and effect of indicated inhibitors on IL-6 secretion. (a) U266 and IM9, multiple myeloma cell lines were starved for 8 h and then treated with or without 1 ng/mL TNF α for 10 min. Each cell line was harvested and prepared for immunoblotting. Upregulation of MAPK, JNK, and PI3K/AKT by TNF α was examined with indicated antibodies. GAPDH was used to ensure equal loading. (b) U266 cells were pretreated with various signal inhibitors for 1 h, respectively. These cells were then stimulated with TNF α for 10 min, and cell supernatants were harvested for ELISA to determine IL-6 concentration induced by TNF α . Bars represent the mean ± SEM from three independent experiments. c: control; 1: TNF α only; 2: TNF α plus 10 μ M PD98059; 3: TNF α plus 20 μ M LY294002; 4: TNF α plus 10 μ M SB203580; 5: TNF α plus 40 nM JNK inhibitor II.

Article Snippet: IM9 was obtained from the Korean Cell Line Bank (Chongro-gu, Seoul, Republic of Korea).

Techniques: Activation Assay, Protein-Protein interactions, Western Blot, Enzyme-linked Immunosorbent Assay, Concentration Assay, Control