Cells were acquired within a week after multimer staining.
Following two washing steps and fixation in PFA, cells were acquired on an LSR Fortessa (BD).
Cells were acquired mycoplasma-free, but have not been specifically tested for mycoplasma in this study.
Stained cells were acquired on a FACSVerse (BD Biosciences, San Diego, CA, USA).
Cells were acquired on a LSRII flow cytometer (BD Biosciences) and analysed using FlowJo software (Treestar, Ashland, OR, USA).
Stained cells were acquired on a BD LSR Fortessa cell analyzer and analyzed with FlowJo (Treestar) software.
Typically, for each condition, at least 30 field of views comprising one or more cells were acquired from a total of at least 3 independent biological replicates.
Z-stacks (~20 images, 1 every micron) of identified cells were acquired every 10 minutes before, during, and after agonist exposure.
A total of 10,000 cells were acquired for each flow cytometric analysis.
Images of the cells were acquired using an LSM-710 confocal microscope (Carl Zeiss, Oberkochen, Germany).
After washing and fixing, at least 104 cells were acquired and analyzed.
