Journal: Heliyon

Article Title: Mesendogen, a novel inhibitor of TRPM6, promotes mesoderm and definitive endoderm differentiation of human embryonic stem cells through alteration of magnesium homeostasis

doi: 10.1016/j.heliyon.2015.e00046

Figure Lengend Snippet: MEG enhances mesoderm differentiation of hiPSCs and of variations of mesoderm induction conditions. (A) FACS analysis of PE and APC in H9 hESCs cultured in mTeSR1 medium. Cells were stained with T-PE and EOMES-APC antibodies (T-PE/EOMES-APC) or non-specific IgG isotype control antibodies conjugated with the same fluorophores (isotype control). Isotype IgG, when used as a gating control, gave rise to a high percentage of false-positive cells in the undifferentiated mTeSR1 culture (98.42% PE + APC + in the T-PE/EOMES-APC stained sample). n > 10 independent experiments. (B) FACS analysis of T and EOMES in MMW2 hiPSCs differentiated in A-BVF, MEG+A-BVF, and MEG->A-BVF conditions for 2 days. Cells were plated at 0.5 × 10 5 cells/cm 2 for A-BVF and MEG->A-BVF conditions and 0.25 × 10 5 cells/cm 2 for MEG+A-BVF condition. Cells cultured in mTeSR1 medium were used as a gating control. n = 2 independent experiments. (C) FACS analysis of T in H1 hESCs differentiated in BMP4 (10 ng/ml), VEGF (10 ng/ml), and bFGF (20 ng/ml) (BVF condition) for 1.5 days, with or without MEG (10 μM) pre-treatment. n = 2 independent experiments. (D) FACS analysis of T in H1 hESCs differentiated in BMP4 (10 ng/ml) and bFGF (20 ng/ml) (BF condition) for 1.5 days, with or without MEG pre-treatment. n > 5 independent experiments.

Article Snippet: Mouse IgG 2B -APC isotype control , BioLegend , 400319 , 1:20 (FACS).

Techniques: Cell Culture, Staining

Journal: Heliyon

Article Title: Mesendogen, a novel inhibitor of TRPM6, promotes mesoderm and definitive endoderm differentiation of human embryonic stem cells through alteration of magnesium homeostasis

doi: 10.1016/j.heliyon.2015.e00046

Figure Lengend Snippet: MEG enhances definitive endoderm differentiation of hiPSCs and of variations of definitive endoderm induction conditions. (A) FACS analysis of SOX17 and FOXA2 in H9 hESCs cultured in mTeSR1 medium. Cells were stained with SOX17-APC and FOXA2-Alexa488 antibodies (SOX17-APC/FOXA2-Alexa488) or non-specific IgG isotype control antibodies conjugated with the same fluorophores (isotype control). Isotype IgG, when used as a gating control, gave rise to a high percentage of false-positive cells in the undifferentiated mTeSR1 culture (∼50% APC + /Alexa488 + and ∼98% Alexa488 + in the SOX17-APC/FOXA2-Alexa-488 stained sample). n > 10 independent experiments. (B) FACS analysis of SOX17 and FOXA2 in MMW2 hiPSCs cells induced to undergo definitive endoderm differentiation under AWS, MEG+AWS, and MEG->AWS conditions for 7 days. Cells were plated at 1.0 × 10 5 cells/cm 2 for AWS and MEG+AWS conditions, and at 0.5 × 10 5 cells/cm 2 for MEG->AWS condition. Cells cultured in mTeSR1 medium were used as a gating control. n = 2 independent experiments. (C) FACS analysis of SOX17 and FOXA2 in H9 hESCs induced to undergo definitive endoderm differentiation for 5 days in AW-B27, MEG+AW-B27, and MEG->AW-B27 conditions. B-27 supplement was used in these conditions instead of serum. Cells were plated at 1.0 × 10 5 cells/cm 2 . n > 3 independent experiments. (D) FACS analysis of SOX17 and FOXA2 in H9 hESCs induced to undergo definitive endoderm differentiation for 5 days in AW-ITS, MEG+AW-ITS, and MEG->AW-ITS conditions. ITS (insulin, transferrin, and selenium) supplement was used in these conditions instead of serum. Cells were plated at 2.0 × 10 5 cells/cm 2 . n = 2 independent experiments.

Article Snippet: Mouse IgG 2B -APC isotype control , BioLegend , 400319 , 1:20 (FACS).

Techniques: Cell Culture, Staining

Journal: Heliyon

Article Title: Mesendogen, a novel inhibitor of TRPM6, promotes mesoderm and definitive endoderm differentiation of human embryonic stem cells through alteration of magnesium homeostasis

doi: 10.1016/j.heliyon.2015.e00046

Figure Lengend Snippet: Antibody sources and dilutions.

Article Snippet: Mouse IgG 2B -APC isotype control , BioLegend , 400319 , 1:20 (FACS).

Techniques:

Journal: AIDS Research and Treatment

Article Title: Expression and Function of the Chemokine, CXCL13, and Its Receptor, CXCR5, in Aids-Associated Non-Hodgkin's Lymphoma

doi: 10.1155/2010/164586

Figure Lengend Snippet: AIDS-NHL cell lines express CXCR5, as shown by flow cytometry. The AIDS-BL cell line, 2F7 (a), and the AIDS-DLBCL cell line, R (b), were stained for CXCR5 expression using an indirect staining protocol, as noted in . First, cells were stained with a rat IgG 2b anti-CXCR5 antibody (dotted lines). As a control, some cells were stained with a rat IgG 2b isotype control antibody (solid lines). Cells were then stained with a PE-conjugated goat anti-rat IgG secondary antibody, and examined by flow cytometry. During the flow cytometry acquisition stage, at least 5,000 cells/events were acquired per tube/condition. During analysis, dead cells were excluded using forward- and side-scatter.

Article Snippet: For CXCR5, a rat antihuman CXCR5 antibody (clone RF8B2, R&D Systems) or IgG 2b isotype control (clone KLH/G2b-1-2, Southern Biotechnology Associates, Birmingham, AL) was used (2 μ g/mL, 2 hours).

Techniques: Flow Cytometry, Staining, Expressing, Control