macstm mouse treg selection kit  (Miltenyi Biotec)

Journal: Apoptosis

Article Title: Regulatory T cells are less sensitive to glucocorticoid hormone induced apoptosis than CD4 + T cells

doi: 10.1007/s10495-020-01629-x

Figure Lengend Snippet: Flow cytometric analysis of the purity of sorted CD4 + CD25 high+ thymocytes and splenocytes after negative selection of CD4 + cells followed by CD25 + positive selection of Treg cells. Foxp3 intracellular staining was used to test the purity of Treg subpopulations. The figure shows representative flow cytometric plots

Article Snippet: In some experiments the CD4 + T cells were subjected to further positive selection to obtain CD4 + CD25 + Treg cells using EasySepTM Mouse Treg Positive Selection Kit (Stemcell Technologies, Vancouver, Canada) following the manufacturer’s instructions.

Techniques: Selection, Staining

Journal: Apoptosis

Article Title: Regulatory T cells are less sensitive to glucocorticoid hormone induced apoptosis than CD4 + T cells

doi: 10.1007/s10495-020-01629-x

Figure Lengend Snippet: a Treg cells express higher level of apoptosis-related proteins than CD4 + T cells. Analysis of 21 apoptosis-related protein expression in the cell lysates of untreated, sorted pTreg cells and CD4 + splenocytes. Bars show the average OD values of each analyte from at least three independent experiments (n = 6). b Effect of DX treatment on the expression of 21 apoptosis-related molecules in T cell subpopulations. Protein arrays were used to detect the in vitro DX treatment (4 h) induced changes of apoptosis-related molecules in cell lysates of sorted CD4 + T cells and pTreg cells. Bars show the DX-induced relative changes (%) of the average OD values of each analyte compared to untreated controls from at least three independent experiments (n = 6). Relative changes were calculated with the following formula: [(OD DX – OD Ctrl )/OD Ctrl ] × 100%

Article Snippet: In some experiments the CD4 + T cells were subjected to further positive selection to obtain CD4 + CD25 + Treg cells using EasySepTM Mouse Treg Positive Selection Kit (Stemcell Technologies, Vancouver, Canada) following the manufacturer’s instructions.

Techniques: Expressing, In Vitro

Journal: Apoptosis

Article Title: Regulatory T cells are less sensitive to glucocorticoid hormone induced apoptosis than CD4 + T cells

doi: 10.1007/s10495-020-01629-x

Figure Lengend Snippet: Effect of DX on basal Ca 2+ levels in tTreg and pTreg cells. Short-term (30 min) DX treatment induced elevation of basal cytosolic free Ca 2+ levels in thymic Treg cells. In both organs, the baseline Ca 2+ levels were higher in Treg cells than in the corresponding CD4 + T cells. The columns show the Fluo 3-AM fluorescence intensities (MFI) of Tregs minus MFI of CD4 + T cells. Data represent mean ± SEM (n = 4). Significant change is indicated with asterisk ( p < 0.05)

Article Snippet: In some experiments the CD4 + T cells were subjected to further positive selection to obtain CD4 + CD25 + Treg cells using EasySepTM Mouse Treg Positive Selection Kit (Stemcell Technologies, Vancouver, Canada) following the manufacturer’s instructions.

Techniques: Fluorescence

Journal: Apoptosis

Article Title: Regulatory T cells are less sensitive to glucocorticoid hormone induced apoptosis than CD4 + T cells

doi: 10.1007/s10495-020-01629-x

Figure Lengend Snippet: Calcium response is low in thymic and splenic Treg cells. Effect of TCR (anti-CD3) stimulation in thymic CD4 SP and tTreg cells was measured in the presence or absence of DX ( a ). Calcium influx kinetics in splenic CD4 + and Treg cells upon specific TCR-mediated activation (anti-CD3) in the presence or absence of DX ( b ). Data were calculated as the relative ratio of calcium values of Fluo 3-AM fluorescence intensities (median). Columns show the relative ratio of peak to baseline calcium flux measured as Fluo 3-AM fluorescence intensities (MFI) ( c ). The graphs show the mean ± SEM of at least three independent experiments (n = 9). Significant changes are indicated with asterisk (* p < 0.05, ***p < 0.001)

Article Snippet: In some experiments the CD4 + T cells were subjected to further positive selection to obtain CD4 + CD25 + Treg cells using EasySepTM Mouse Treg Positive Selection Kit (Stemcell Technologies, Vancouver, Canada) following the manufacturer’s instructions.

Techniques: Activation Assay, Fluorescence