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: Oncotarget

Article Title: Long-term treatment with imatinib results in profound mast cell deficiency in Ph+ chronic myeloid leukemia

doi:

Figure Lengend Snippet: A: Isolated cord blood MNC (1×10 6 /ml) were cultured in 24-well plates in RPMI medium containing 10% FCS, IL-6 and SCF (each 100 ng/ml) in the presence or absence (CO) of various concentrations of imatinib (0.001-1 μM) at 37°C for 28 days. Thereafter, the total numbers of mast cells (MC) per well were determined by measuring total cell counts and the percentage of MC on Giemsa-stained slides. Cells were counted using an Olympus AX-1 microscope equipped with a 100x/1.35 UPlan-Apo objective lens. Results represent the mean±S.D. from 3 independent experiments. Asterisk: p<0.05. B: Total histamine levels per well assessed by RIA (left panel) and total tryptase levels assessed by FEIA (right panel) were determined on day 28. Results represent the mean±S.D. from 3 independent experiments. Asterisk: p<0.05. C: Expression of tryptase mRNA levels (left panel) and KIT (CD117) mRNA levels (right panel) determined by qPCR on day 28. Results show tryptase and KIT mRNA expression levels as percent of ABL mRNA levels, and represent the mean±S.D. from 3 independent experiments. Asterisk: p<0.05.

Article Snippet: Recombinant human (rh) stem cell factor (SCF) was from Strathmann Biotech (Hannover, Germany) and rh interleukin-6 (IL-6) from Novartis Pharma AG.

Techniques: Isolation, Cell Culture, Staining, Microscopy, Expressing

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