Splenocytes and siLPLs were incubated with a rat anti-mouse CD16/CD32 mAb (BioLegend, USA) for 30 min to block Fc receptors. Then, the cells were stained on ice with fluorochrome-conjugated monoclonal antibodies for 30 min as previously described [24 (link)]. To perform intracellular staining, cells were treated using a Fixation/Permeabilization Solution kit (BD Pharmingen™, USA) and then stained with the appropriate antibodies. The antibodies used in this study are listed in Additional file 2 : Figure S6. Cells were assayed with a Symphony A5 cytometer (BD Biosciences), and the resulting data was analysed using FlowJo (ver. 10.4).
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