Cyto x cytoperm plus fixation permeabilization kit

Mouse lymphocytes were stained with following uorochrome conjugated antibodies: CD4-PerCP Cy5.5, CD25-APC, Foxp3-PE, IFN-γ-APC, IL-17-FITC, B220-APC, CD19-PerCP, CD1d-PE, CD5-FITC, IL-10-APC, CD138-PE, and GL7-FITC. Human lymphocytes were stained with following uorochrome conjugated antibodies: CD4-PECy7, CD25-APC, Foxp3-PE, IFN-γ-APC, IL-17-FITC,, CD19-FITC, IL-10-APC, CD138-PB450 Before intracellular staining, cells were stimulated for 4 h with phorbol myristate acetate (25 ng/mL) and ionomycin (250 ng/mL) in the presence of Golgistop (BD Biosciences, San Jose, CA, USA). Intracellular staining was xed using a BD Cyto x/ Cytoperm Plus Fixation/Permeabilization Kit and BD Golgistop Kit (BD Biosciences, San Jose, CA). Foxp3, transcription factor was xed using a Foxp3/Transcription Factor Staining buffer set (eBioscience, San Diego, CA) following the manufacturer's instructions. Flow cytometric analysis was performed using a cytoFLEX Flow Cytometer (Beckman Coulter, Brea, CA, USA) and collected data were analyzed using the FlowJo software (Tree Star, Ashland, OR).

Mouse lymphocytes were stained with following uorochrome conjugated antibodies: CD4-PerCP Cy5.5, CD25-APC, Foxp3-PE, IFN-g-APC, IL-17-FITC, B220-APC, CD19-PerCP, CD1d-PE, CD5-FITC, IL-10-APC, CD138-PE, and GL7-FITC. Human lymphocytes were stained with following uorochrome conjugated antibodies: CD4-PECy7, CD25-APC, Foxp3-PE, IFN-g-APC, IL-17-FITC, , CD19-FITC, IL-10-APC, CD138-PB450 Before intracellular staining, cells were stimulated for 4 h with phorbol myristate acetate (25 ng/mL) and ionomycin (250 ng/mL) in the presence of Golgistop (BD Biosciences, San Jose, CA, USA). Intracellular staining was xed using a BD Cyto x/ Cytoperm Plus Fixation/Permeabilization Kit and BD Golgistop Kit (BD Biosciences, San Jose, CA). Foxp3, transcription factor was xed using a Foxp3/Transcription Factor Staining buffer set (eBioscience, San Diego, CA) following the manufacturer's instructions. Flow cytometric analysis was performed using a cytoFLEX Flow Cytometer (Beckman Coulter, Brea, CA, USA) and collected data were analyzed using the FlowJo software (Tree Star, Ashland, OR).

Mouse lymphocytes were stained with following uorochrome conjugated antibodies: CD4(L3T4)-PerCP Cy5.5, CD25(PC61)-APC, Foxp3(FJK-16s)-PE, IFN-g(XMG1.2)-APC, IL-17(eBio17B7)-FITC, B220(RA3-6B2)-APC, CD19(eBio1D3(1D3))-PerCP, CD1d(1B1)-PE, CD5(53-7.3)-FITC, IL-10(JES5-16E3)-APC, CD138(281-2)-PE, and T-and B-Cell Activation Antigen (GL7)-FITC. Human lymphocytes were stained with following uorochrome conjugated antibodies: CD4(RPA-T4)-PECy7, CD25(BC96)-APC, Foxp3(259D/C7)-PE, IFNg(4S.B3)-APC, IL-17(eBio64DEC1-FITC, , CD19(HIB19)-FITC, IL-10(JES3-19F1)-APC, CD138(MI15)-PB450. Before intracellular staining, cells were stimulated for 4 h with phorbol myristate acetate (25 ng/mL) and ionomycin (250 ng/mL) in the presence of Golgistop (BD Biosciences, San Jose, CA, USA). Intracellular staining was xed using a BD Cyto x/ Cytoperm Plus Fixation/Permeabilization Kit and BD Golgistop Kit (BD Biosciences, San Jose, CA). Foxp3, transcription factor was xed using a Foxp3/Transcription Factor Staining buffer set (eBioscience, San Diego, CA) following the manufacturer's instructions. Flow cytometric analysis was performed using a cytoFLEX Flow Cytometer (Beckman Coulter, Brea, CA, USA) and collected data were analyzed using the FlowJo software (Tree Star, Ashland, OR).