Apc cy7 α cd19 1d3

PECs were collected and washed once. For latency experiments, cells were counted and CD11b+ cells were selected by MACS column purification using the CD11b MicroBead kit and LS columns (Miltenyi Biotec). Total PECs were used for MHV68 acute infection experiments. CD11b+ cells or whole PECs were Fc blocked with α-CD16/32 (Biolegend) and stained with Alexafluor 647-α-CD102 (3C4 (MIC2/4), Biolegend), APC-Cy7-α-CD19 (1D3, BD Biosciences), redFluor 710-α-CD11b (M1/70, Tonbo) or APC-Cy7-α-CD11b (M1/70, Biolegend), and R718-α-CD102 (3C4 (MIC2/4), BD Biosciences). Cells were counted again and resuspended in 3 x 10⁶ cells/mL before adding CC4F-AM (LiveBLAzer FRET-B/G Loading Kit with CCF4-AM, ThermoFisher Scientific). Cells were incubated for 1 hour, then washed with PBS and fixed with 2% formaldehyde. Samples were immediately run on a Novocyte 3000 (ACEA Biosciences).

PECs were collected and washed once. For latency experiments, cells were counted and CD11b+ cells were selected by MACS column purification using the CD11b MicroBead kit and LS columns (Miltenyi Biotec). Total PECs were used for MHV68 acute infection experiments. CD11b+ cells or whole PECs were Fc blocked with α-CD16/32 (Biolegend) and stained with Alexafluor 647-α-CD102 (3C4 (MIC2/4), Biolegend), APC-Cy7-α-CD19 (1D3, BD Biosciences), redFluor 710-α-CD11b (M1/70, Tonbo) or APC-Cy7-α-CD11b (M1/70, Biolegend), and R718-α-CD102 (3C4 (MIC2/4), BD Biosciences). Cells were counted again and resuspended in 3 x 10⁶ cells/mL before adding CC4F-AM (LiveBLAzer FRET-B/G Loading Kit with CCF4-AM, ThermoFisher Scientific). Cells were incubated for 1 hour, then washed with PBS and fixed with 2% formaldehyde. Samples were immediately run on a Novocyte 3000 (ACEA Biosciences).