Percp cy5.5 anti gr 1

RBC-depleted spleen cell suspensions prepared from WT and TAX1BP1^−/−mouse were labeled with CD11b and CD90.2 microbeads (Miltenyl Biotec). Then the B cells were enriched by negative selection using autoMACSpro separator (Miltenyl Biotec) according to the manufacturer’s instructions. The collected cells were stained with the following antibodies: APC anti-B220, PE anti-Fas, PE anti-CD138 (Syndecan-1), FITC anti mouse IgM, FITC anti-GL7 antigen (BioLegend) and 7-AAD (BioLegend). Single-cell suspensions of bone marrow from WT and TAX1BP1^−/− mouse were stained with following antibodies: PE/Cy7 anti-TCR β chain, 7-AAD, PerCP-Cy5.5 anti-Gr-1, APC anti-B220, and FITC anti-mouse IgM (BioLegend). Single-cell suspensions of total splenocytes from WT and TAX1BP1^−/− mouse were stained with different combinations of the following antibodies: PE/Cy7 anti-TCR β chain, PerCP-Cy5.5 anti-Gr-1, 7-AAD, FITC anti-mouse IgM, APC anti-CD23, PE anti-CD21 (BioLegend), or PerCP-Cy5.5 anti-Gr-1, 7-AAD, PE/Cy7 anti-B220, PE anti-CD93, FITC anti-mouse IgM and APC-CD23 (BioLegend). WT DT40 cells, TAX1BP1^−/−/− cells and TAX1BP1^−/−/−/Tax1bp1 cells were stained with FITC conjugated anti chicken IgM (Bethyl). FACS Verse (BD) and SH800 (Sony Biotechnology Inc.) were used for flow cytometry, and results were analyzed with FlowJo software.