Live dead zombie

All antibodies were purchased from BD Pharmingen, eBioscience, Invitrogen, BioLegend, and the UCSF hybridoma core, or were produced in the Krummel laboratory. For surface staining, cells were incubated with anti-Fc receptor antibody (clone 2.4G2) and then stained with antibodies in PBS + 2% FCS for 30 min on ice. Viability was assessed by staining with fixable Live/Dead Zombie (BioLegend) or DAPI. Flow cytometry was performed on a BD Fortessa instrument. Analysis of flow cytometry data was done using FlowJo (Treestar) software. Cell sorting was performed using a BD FACS Aria II.

All antibodies were purchased from BD Pharmingen, eBioscience, Invitrogen, Biolegend, the UCSF hybridoma core, or were produced in the Krummel Lab. For surface staining cells were incubated with anti-Fc receptor antibody (clone 2.4G2) and stained with antibodies in PBS + 2 % FCS for 30 min on ice. Viability was assed by staining with fixable Live/Dead Zombie (Biolegend) or DAPI. For intracellular staining, mice were injected with 10 ug/gram of body weight with Brefeldin A (Cayman) 6 hr prior to harvest, cells were stained with antibodies against surface markers, then fixed with 2 % PFA for 10 min at 25 °C and permeabilized with 0.2 % Saponin then stained with target antibody. All flow cytometry was performed on a BD Fortessa flow cytometer. Analysis of flow cytometry data was done using Flowjo (Treestar). Cell sorting was performed using a BD FACS Aria II.

All antibodies were purchased from BD Pharmingen, eBioscience, Invitrogen, Biolegend, the UCSF hybridoma core, or were produced in the Krummel Lab. For surface staining cells were incubated with anti-Fc receptor antibody (clone 2.4G2) and stained with antibodies in PBS + 2% FCS for 30 min on ice. Viability was assessed by staining with fixable Live/Dead Zombie (Biolegend) or DAPI. All flow cytometry was performed on a BD Fortessa flow cytometer. Analysis of flow cytometry data was done using Flowjo (Treestar).