Percp cyanine5.5 anti mouse cd14 antibody

The isolated murine splenocytes were incubated with B. burgdorferi (1 × 10⁶ cells/mL) alone or mixture with 1 μg/mL rIsC1ql3 for 6 h. The inhibitors of intracellular protein transport, monensin (Invitrogen, #00-4505-51) and brefeldin A (Invitrogen, #00-4506-51) were added during the last hours of in vitro activation. The cells were then incubated with Fc receptor antibody (TruStain FcX^™ anti-mouse CD16/32) (BioLegend, #101320), and the antibodies of FITC anti-mouse CD3 antibody (BioLegend, #100204), APC anti-mouse CD4 antibody (BioLegend, #100412), and Brilliant Violet 711^™ anti-mouse CD8a antibody (BioLegend, #100747), PerCP/Cyanine5.5 anti-mouse CD14 antibody (BioLegend, #123313), PE anti-mouse NK-1.1 antibody (BioLegend, #108708), and LIVE/DEAD^™ fixable violet stain kit (Invitrogen, #L34955) on ice for 20 min. Then the cells were fixed and permeabilized using BD Cytofix/Cytoperm fixation/permeabilization solution (BD Biosciences, #555028). Cells were washed and stained by intracellular staining for 30 min at 4°C using APC/Cyanine7 anti-mouse IFN-γ antibody (BioLegend, #505849). The samples were run through BD LSRII (BD bioscience). The data was then analyzed by FlowJo.