Journal: PLoS ONE

Article Title: Curcumin Inhibits CD4 + T Cell Activation, but Augments CD69 Expression and TGF-β1-Mediated Generation of Regulatory T Cells at Late Phase

doi: 10.1371/journal.pone.0062300

Figure Lengend Snippet: CD4 + T cells were cultured in the presence of anti-CD2/CD3/CD28 antibody-coated beads only (Act.) or with 2 µg/mL curcumin (Cur.) for 3 days. Act. (1/2) and Act. (1/10) indicate a 1∶2 and 1∶10 bead-to-cell ratio, respectively. (A) Cells were harvested at the indicated time point and the percentage of CD69 + cells was determined by flow cytometric analysis. (B–D) After 3 days of culture, cells were harvested, washed with PBS, and then transferred to a new cell culture plate in fresh media for an additional 3 days. The cells were then stained and analyzed by flow cytometry. (B) Histograms of CD69 expression and the total percentage of CD69 + cells. (C) The numbers in each plot and the number in blanket indicate the percentage of cells in each respective area and the percentage of CD69 + cells among cells positive with the Y axis, respectively. (D) The percentage of total cells positive for each molecule. Data are representative of 3 replicate experiments yielding similar results. (A, B and D) Data are presented as the mean ± SD. * P <0.05, *** P <0.001.

Article Snippet: Fluorophore-conjugated monoclonal antibodies for surface or intracellular molecules were purchased from BD Bioscience (San Jose, CA, USA), unless otherwise stated, as follows; anti-Annexin-V FITC, -CD25 APC or PE, -CD69 APC or PE, -CD45RO PE, -CD27 FITC, -CCR7 Alexa647, -IL-12RβI PE, -L-selectin PE, -integrin β7 PE, -CD40L PE, -TGF-β PE (R&D Systems, Minneapolis, MN, USA), -IFN-γ APC, -TNF-α PE, -IL-10 PE, -IL-13 PE, -IL-17A APC, -Foxp3 PE antibodies.

Techniques: Cell Culture, Staining, Flow Cytometry, Expressing

Journal: PLoS ONE

Article Title: Curcumin Inhibits CD4 + T Cell Activation, but Augments CD69 Expression and TGF-β1-Mediated Generation of Regulatory T Cells at Late Phase

doi: 10.1371/journal.pone.0062300

Figure Lengend Snippet: (A, B) CD4 + T cells were cultured in the presence of anti-CD2/CD3/CD28 antibody-coated beads only (Act.; 1∶10 for bead to cell) or with 2 µg/mL curcumin (Cur.) for 3 days. Cells were then harvested and the percentage of CD69 + cells was determined by flow cytometric analysis. (A) After 48 hours of culture, cells were treated with an additional 2 µg/mL of curcumin (Cur. 48 hours). The number in each panel indicates the total percentage of CD69 + cells. The results are the representative of 3 replicate experiments yielding similar results. (B) After 48 hours of culture, cells were treated with 10 µM of U0126 (ERK inhibitor), SP600125 (JNK inhibitor), or SB203580 (p38 MAPK inhibitor). (C) CD4 + T cells were cultured in the presence of anti-CD2/CD3/CD28 antibody-coated beads (Act.; 1∶10 for bead-to-cell ratio) for 3 days, with 10 µM of U0126 (ERK inhibitor), SP600125 (JNK inhibitor), or SB203580 (p38 MAPK inhibitor) added 1 hour prior to treatment with an additional 2 µg/mL of curcumin (Cur. 48hrs) after 48 hours of culture. CD69 expression was assessed by flow cytometry. Data are presented as the mean ± SD. ** P <0.01, *** P <0.001.

Article Snippet: Fluorophore-conjugated monoclonal antibodies for surface or intracellular molecules were purchased from BD Bioscience (San Jose, CA, USA), unless otherwise stated, as follows; anti-Annexin-V FITC, -CD25 APC or PE, -CD69 APC or PE, -CD45RO PE, -CD27 FITC, -CCR7 Alexa647, -IL-12RβI PE, -L-selectin PE, -integrin β7 PE, -CD40L PE, -TGF-β PE (R&D Systems, Minneapolis, MN, USA), -IFN-γ APC, -TNF-α PE, -IL-10 PE, -IL-13 PE, -IL-17A APC, -Foxp3 PE antibodies.

Techniques: Cell Culture, Expressing, Flow Cytometry

Journal: Molecular Oncology

Article Title: Single‐cell protein profiling defines cell populations associated with triple‐negative breast cancer aggressiveness

doi: 10.1002/1878-0261.13365

Figure Lengend Snippet: Overview of antibodies and reagents used in study.

Article Snippet: CD69 , FN50 , 144Nd , Fluidigm , 3144018B.

Techniques: Cytometry, Marker