Flowjo 7.5.5 Download |BEST|

Flowjo 7.5.5 Download |BEST|

Flowjo 7.5.5 Download ::: https://ssurll.com/2t7AWX

T cells were seeded onto flat bottom polystyrene 48-well plates (Costar, Corning Inc.) at 106 cells/ml in a volume of 500 μl per well and stimulated as described above. After 16 h, cells were stained with FITC- or PE-labeled mAbs against CD25 and CD69 (BD Pharmingen, BioLegend) and analyzed by flow cytometry using a FACSCalibur and CellQuest software or BD LSRFortessa and FACSDiva Software 6.1.3 (all from BD Biosciences), and FlowJo 7.5.5 (Tree Star, Inc.).

T cells were centrifuged, resuspended in 1 ml of PBS Dulbecco (Biochrom AG) and labeled with 2,5 μM CFSE (Invitrogen) for 15 min at 37°C. Then cells were washed twice, resuspended in medium at the density of 106 cells/ml, seeded onto flat bottom polystyrene 48-well plates (Costar, Corning Inc.) in a volume of 500 μl/well and stimulated as described above. Cells were then cultured for 72 h and proliferation was assessed by CFSE dilution using FACSCalibur and CellQuest software or BD LSRFortessa and FACSDiva Software 6.1.3 (all from BD Biosciences), and FlowJo 7.5.5 (Tree Star, Inc.).

Cell-cycle fractions were determined by propidium iodide nuclear staining. Briefly, cells were harvested, washed in PBS, fixed in 70% ethanol for 30 minutes on ice, and incubated in propidium iodide solution (20 μg/mL propidium iodide, 0.2 mg/mL RNAse A in PBS) for 30 minutes at 37°C. Data were collected on a FACSCalibur flow cytometer (BD Biosciences) and analyzed with FlowJo Version 7.5.5 software (TreeStar). Results represent the mean value of 3 independent experiments. 2b1af7f3a8