Journal: Molecular Medicine Reports

Article Title: TNF-α enhances vascular cell adhesion molecule-1 expression in human bone marrow mesenchymal stem cells via the NF-κB, ERK and JNK signaling pathways

doi: 10.3892/mmr.2016.5314

Figure Lengend Snippet: Effect of TNF-α on VCAM-1 expression in MSCs. MSCs were stimulated with TNF-α (0, 6.25, 12.50, 25, 50 and 100 ng/ml) for 24 h and ana-lyzed by flow cytometry. Data were obtained from three separate experiments and are presented as means ± standard deviation. * P<0.05 vs. control, # P<0.05 vs. 50 ng/ml. VCAM-1, vascular cell adhesion molecule 1; TNF-α, tumor necrosis factor-α; MSCs,mesenchymal stem cells.

Article Snippet: MSCs were divided into four groups: i) Without intervention; ii) stimulant alone, 50 ng/ml TNF-α (ProSpec-Tany TechnoGene, Ltd., East Brunswick, NJ, USA); iii) inhibitors alone, 10 µ M ERK inhibitor, U0126 (Cell Signaling Technology, Inc., Danvers, MA, USA), 1 mM NF-κB inhibitor, pyrrolidine dithiocarbamate (PDTC; Merck Millipore, Darmstadt, Germany) or 20 µ M JNK inhibitor, SP600125 (Merck Millipore); and iv) inhibitor + stimulant, inhibitor stimulation (as above) followed by 50 ng/ml TNF-α.

Techniques: Expressing, Flow Cytometry, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: TNF-α enhances vascular cell adhesion molecule-1 expression in human bone marrow mesenchymal stem cells via the NF-κB, ERK and JNK signaling pathways

doi: 10.3892/mmr.2016.5314

Figure Lengend Snippet: PDTC, U0126, and SP600125 blocked TNF-α-induced VCAM-1 expression in MSCs. (A) The appropriate active MSCs were analyzed by flow cytometry. (B) VCAM-1 expression of MSCs in control cells measured by flow cytometry. The effects of (C) TNF-α or (D) nuclear factor-κB inhibitor, PDTC, (E) ERK inhibitor, U0126, (F) JNK inhibitor, SP600125 on VCAM-1 expression levels were measured by flow cytometry. Additionally, the combined effect of TNF-α and (G) PDTC, (H) U0126 and (I) SP600125 on VCAM-1 expression were determined by flow cytometry and (J) quantified. (K) Reverse transcription-quantitative polymerase chain reaction was performed to measure the VCAM-1 mRNA expression levels from the same treatment groups. Data were obtained from three independent experiments and are presented as means ± standard deviation. * P<0.05 vs. control, ** P<0.05 vs. TNF-α group. PDTC, pyrrolidine dithiocarbamate; TNF-α, tumor necrosis factor-α; VCAM-1, vascular cell adhesion molecule 1; ERK, extracellular signal-regulated kinase; JNK, c-Jun N-terminal kinase.

Article Snippet: MSCs were divided into four groups: i) Without intervention; ii) stimulant alone, 50 ng/ml TNF-α (ProSpec-Tany TechnoGene, Ltd., East Brunswick, NJ, USA); iii) inhibitors alone, 10 µ M ERK inhibitor, U0126 (Cell Signaling Technology, Inc., Danvers, MA, USA), 1 mM NF-κB inhibitor, pyrrolidine dithiocarbamate (PDTC; Merck Millipore, Darmstadt, Germany) or 20 µ M JNK inhibitor, SP600125 (Merck Millipore); and iv) inhibitor + stimulant, inhibitor stimulation (as above) followed by 50 ng/ml TNF-α.

Techniques: Expressing, Flow Cytometry, Real-time Polymerase Chain Reaction, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: TNF-α enhances vascular cell adhesion molecule-1 expression in human bone marrow mesenchymal stem cells via the NF-κB, ERK and JNK signaling pathways

doi: 10.3892/mmr.2016.5314

Figure Lengend Snippet: Activity of NF-κB, ERK, or JNK signaling in MSCs treated with TNF-α + inhibitors and TNF-α alone. (A) Effect of TNF-α and NF-κB inhibitor, PDTC on NF-κB signaling. (B) Effect of TNF-α and ERK inhibitor, U0126 on ERK signaling. (C) Effect of TNF-α and JNK inhibitor, SP600125 on the JNK signaling pathway. Data were obtained from three independent experiments and are presented as means ± standard deviation. * P<0.05 vs. control, ** P<0.05 vs. TNF-α group. p, phosphorylated; NF-κB, nuclear factor-κB; ERK, extracellular signal-regulated kinase; JNK, c-Jun N-terminal kinase; TNF-α, tumor necrosis factor-α; MSCs, mesenchymal stem cells.

Article Snippet: MSCs were divided into four groups: i) Without intervention; ii) stimulant alone, 50 ng/ml TNF-α (ProSpec-Tany TechnoGene, Ltd., East Brunswick, NJ, USA); iii) inhibitors alone, 10 µ M ERK inhibitor, U0126 (Cell Signaling Technology, Inc., Danvers, MA, USA), 1 mM NF-κB inhibitor, pyrrolidine dithiocarbamate (PDTC; Merck Millipore, Darmstadt, Germany) or 20 µ M JNK inhibitor, SP600125 (Merck Millipore); and iv) inhibitor + stimulant, inhibitor stimulation (as above) followed by 50 ng/ml TNF-α.

Techniques: Activity Assay, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: TNF-α enhances vascular cell adhesion molecule-1 expression in human bone marrow mesenchymal stem cells via the NF-κB, ERK and JNK signaling pathways

doi: 10.3892/mmr.2016.5314

Figure Lengend Snippet: Effects of TNF-α, PDTC, U0126 and SP600125 on MSC adhesion to human umbilical vein endothelial cells. (A) MSCs were divided into four groups: Untreated control cells; 50 ng/ml TNF-α treatment; 50 ng/ml TNF-α treatment + 1 mM PDTC, 50 ng/ml TNF-α treatment + 10 µ M U0126 or 50 ng/ml TNF-α treatment + 20 mM SP600125. Representative photomicrographs of MSCs are presented (scale bar, 100 µ m). Experiment was performed 4 times. (B) Quantitative analyses of the number of adherent cells. Data are presented as the mean ± standard deviation. * P<0.05 vs. control. TNF-α, tumor necrosis factor-α; PDTC, pyrrolidine dithiocarbamate; MSCs, mesenchymal stem cells.

Article Snippet: MSCs were divided into four groups: i) Without intervention; ii) stimulant alone, 50 ng/ml TNF-α (ProSpec-Tany TechnoGene, Ltd., East Brunswick, NJ, USA); iii) inhibitors alone, 10 µ M ERK inhibitor, U0126 (Cell Signaling Technology, Inc., Danvers, MA, USA), 1 mM NF-κB inhibitor, pyrrolidine dithiocarbamate (PDTC; Merck Millipore, Darmstadt, Germany) or 20 µ M JNK inhibitor, SP600125 (Merck Millipore); and iv) inhibitor + stimulant, inhibitor stimulation (as above) followed by 50 ng/ml TNF-α.

Techniques: Standard Deviation

Journal: Biomedicines

Article Title: Effect of MNCQQ Cells on Migration of Human Dermal Fibroblast in Diabetic Condition

doi: 10.3390/biomedicines10102544

Figure Lengend Snippet: Contents of the QQ Culture Medium.

Article Snippet: rh IL-6 , Peprotec #200-06 , 20 ng/mL.

Techniques: Concentration Assay

Journal: PLoS ONE

Article Title: Generation of Megakaryocytic Progenitors from Human Embryonic Stem Cells in a Feeder- and Serum-Free Medium

doi: 10.1371/journal.pone.0055530

Figure Lengend Snippet: Differentiating hESCs were cultured for 10 days (d) in BMP4, VEGF, SCF and FGF2 and then for a further 3 or 10 d in TPO, SCF and IL-3. Cells were dissociated at d13 or d20 of culture and stained with antibodies directed against CD41 and a panel of hematopoietic markers. A) Gating strategy identifying CD41 + cells at d13 and B) CD41 lo and CD41 + cells at d20 of hESC differentiation. C) Percent expression of hematopoietic - CD45, CD34, CD43; megakaryocytic – CD110, CD42b, CD61, myeloid – CD33 and hemato/endothelial – CD117 and KDR surface markers on the CD41 expressing cells. (Representative FACS panels stained with isotype controls for CD41 staining are shown in ).

Article Snippet: After 10 days, 72 EBs were transferred to each well of a 6 well flat-bottom tissue culture plate containing 7.2 ml of BPEL supplemented with 20 ng/ml rh thrombopoietin (TPO), 25 ng/ml rhSCF and 25 ng/ml rh interleukin (IL)-3 (Peprotech).

Techniques: Cell Culture, Staining, Expressing

Journal: PLoS ONE

Article Title: Mapping the Homodimer Interface of an Optimized, Artificial, Transmembrane Protein Activator of the Human Erythropoietin Receptor

doi: 10.1371/journal.pone.0095593

Figure Lengend Snippet: ( A ) Extracts were prepared from parental BaF3 cells or BaF3/HA-hEPOR cells expressing empty RVY-puro vector (V), EBC5-16, or TC2-3. Where indicated, cells were acutely stimulated with EPO. Samples were immunoprecipitated with anti-HA (3F10) antibody and immunoblotted with anti-phosphotyrosine antibody. Size of protein markers (in kDa) is shown on left. ( B ) Extracts from BaF3/HA-hEPOR cells expressing RVY-puro vector (V), EBC5-16, or TC2-3. Where indicated, cells were acutely stimulated with EPO or RPMI-IL-3 medium. Samples were immunoblotted for phosphorylated JAK2. Blot was reprobed for total JAK2. Size of protein markers (in kDa) is shown on left. ( C ) Extracts from the cells described in ( B ) were immunoblotted for phosphorylated STAT5. Blot was reprobed for total STAT5. Size of protein markers (in kDa) is shown on left.

Article Snippet: For erythroid colony forming assays in methylcellulose, 1×10 4 GFP + -infected cells/mL were washed in Iscove’s Modified Dulbecco’s Medium (L-glutamine, 25 mM HEPES, 3.024 g/L Na 2 CO 3 ) (Gibco) plus 2% FBS (Stem Cell Technologies) and diluted 1∶10 in methylcellulose medium (Methocult H4531, Stem Cell Technologies) containing 20 ng/mL rh-IL-3, 20 ng/mL rh-IL-6 (ConnStem), 50 ng/mL rh-SCF in the presence or absence of 3 U/mL of EPO.

Techniques: Expressing, Plasmid Preparation, Immunoprecipitation