Flow Cytometry
Red blood cells were lysed using ACK buffer (Thermo Fisher). Unspecific binding was blocked by using mouse Fc gamma block (BD Bioscience). Basophils were marked with APC anti-mouse CD49b (clone HMα2, Biolegend) and PE anti-mouse IgE (clone RME-1, Biolegend). Antibody stainings were carried out at 4°C for 30min. After the staining, the cells were washed 3x with PBS. Flow cytometry was performed with CytoFLEX S 4L 13C (B2-R3-V4-Y4) plus 96 DW plate loader (Beckman Coulter Life Sciences, CA, USA) and analyzed using FLOWJO software (TreeStar Inc, Ashland, OR, USA).