Journal: OncoTargets and therapy

Article Title: The roles of serum CXCL16 in circulating Tregs and gastrointestinal stromal tumor cells

doi: 10.2147/OTT.S105245

Figure Lengend Snippet: CXCL16, CXCR6, sCXCL16, and circulating Tregs in GIST specimens. Notes: ( A ) Western blot analysis for CXCL16 and CXCR6 expression in specimens. β-actin was used as an internal loading control. ( B ) Representative results of GIST cancer tissues and corresponding noncancerous tissue by immunohistochemical staining. The nuclei were counterstained with hematoxylin. ( C ) sCXCL16 level in serum of the GIST patients was detected by using ELISA. ( D ) Circulating Tregs were determined by using fluorescence-activated cell sorting. ( E ) Positive correlation between sCXCL16 and Tregs in serum of the GIST patients. Abbreviations: GIST, gastrointestinal stromal tumors; ELISA, enzyme-linked immunosorbent assay; N, normal; C, cancer; sCXCL16, serum CXCL16.

Article Snippet: Serum samples were obtained by centrifugation at 4°C and stored at −80°C for future use. sCXCL16 levels were determined using the enzyme-linked immunosorbent assay (ELISA) method (Human ELISA kit; R & D Systems, Inc.).

Techniques: Western Blot, Expressing, Immunohistochemistry, Staining, Enzyme-linked Immunosorbent Assay, Fluorescence, FACS

Journal: Stem Cells (Dayton, Ohio)

Article Title: TAFA2 Induces Skeletal (Stromal) Stem Cell Migration Through Activation of Rac1‐p38 Signaling

doi: 10.1002/stem.2955

Figure Lengend Snippet: TAFA2 levels are regulated during the process of fracture healing. ( A): Immunohistochemical analysis of TAFA2 protein expression in a fractured femur, 48 hours after induction of fracture. Scale bar: 1 mm. (B): Analysis of TAFA2 immunoreactivity at the fracture site (boxed area from A ) at different time points (2, 24, and 48 hours, 4 days, and 1 week) after induction of femoral fracture (upper panel) and nonfractured femur (lower panel). Scale bar: 100 μm. (C): Quantitative reverse transcription polymerase chain reaction (qRT‐PCR) analysis of TAFA2 mRNA expression in nonfractured and fractured mouse femur at different time points after fracture induction (2 hours, day 1, day 4, and day 14). (D): Flow cytometry analysis of TAFA2 protein expression in bone marrow cells obtained from fractured femur at different time points. Data represent MFI ± SEM from three independent experiments. **, p ≤ .01; two‐tailed unpaired Student t test. (E): qRT‐PCR analysis of IL‐1β mRNA expression in mouse femur before fracture and at different time points after fracture induction (2 hours, day 1, day 4, and day 14). (F): qRT‐PCR analysis of TAFA2 mRNA expression in human skeletal (mesenchymal, stromal) stem cell (hMSC) cultures in the presence of different doses of IL‐1β (1, 10, and 50 ng/ml) for 48 hours. Data represent three independent experiments ± SEM; **, p ≤ .01, two‐tailed paired Student t test. (G, H): Western blot analysis and quantification of TAFA2 protein levels in hMSC cultures in the presence of different doses of IL‐1β. Data represent mean ± SEM from four independent experiments. *, p ≤ .05, two‐tailed unpaired Student t test. (I, J): Flow cytometry analysis of TAFA2 protein expression and percentage of leukocytes (CD45 + ), monocytes (CD14 + ), T cells (CD3 + ), and B cells (CD19 + ) at 1 and 3 days postfracture. Data are obtained from pooled bone marrow cells isolated from fractured femurs of three mice. (K): Enzyme‐linked immunosorbent assay measurement of TAFA2 levels in consecutive serum samples from patients with hip fracture. **, p ≤ .01, two‐tailed paired Student t test. (L): Proposed model for role of TAFA2 in recruiting MSC to the site of fracture. Abbreviations: IL, interleukins; MFI, mean fluorescence intensity; MSC, skeletal (mesenchymal) stem cell.

Article Snippet: ELISA Measurement of TAFA2 Concentrations in Clinical SamplesELISA measurements of TAFA2 concentrations in human serum samples were performed using an established ELISA procedure (Mybiosource, San Diego, USA) according to the manufacturer's instructions.

Techniques: Immunohistochemistry, Expressing, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Flow Cytometry, Two Tailed Test, Western Blot, Isolation, Mouse Assay, Enzyme-linked Immunosorbent Assay, Fluorescence

Journal: American Journal of Respiratory Cell and Molecular Biology

Article Title: Effect of IL-10 on Neutrophil Recruitment and Survival after Pseudomonas aeruginosa Challenge

doi: 10.1165/rcmb.2008-0202OC

Figure Lengend Snippet: Mean (±SEM) lung keratinocyte–derived chemokine (KC), TNF-α, and IL-10 levels 6 and 24 hours after intratracheal administration of P. aeruginosa . In ( A ) and ( B ), mice were infected with P. aeruginosa at a dose of 1.1 × 10 6 CFU. BALFs were collected at 6 hours after infection and measured for KC and TNF-α expression by ELISA with determinations made in triplicate. In ( C ) and ( D ), BALFs were collected at 24 hours after infection and measured for KC and TNF-α expression. ( E ) Endogenous IL-10 (mouse IL-10) and exogenous IL-10 (human IL-10) levels were measured by ELISA in a similar manner from 6- and 24-hour BALF samples. * P

Article Snippet: Lung KC, TNF-α, mouse IL-10, and human IL-10 levels were measured by the ELISA method (Cytoset; Biosource, Invitrogen, Camarillo, CA) according to the manufacturer's instructions.

Techniques: Derivative Assay, Mouse Assay, Infection, Expressing, Enzyme-linked Immunosorbent Assay