4.5. Inflammasome formation assays

For stimulation with fusokine or cytokines, DC media was removed and blank RMPI containing the necessary cytokine was added. Fusokine stimulation on DCs was carried out as described in (Hsieh et al., 2015). DCs were incubated with fusokine/cytokine for 3 h and then activated with lipopolysaccharide (1 μg/mL) overnight. Before collection for experiments, DCs were also primed with ATP (1 μg/mL) 10 min prior to collection. Once collected, activated DCs were lysed and RNA was collected (Quiagen RNEasy) for QPCR as per standard protocols (Applied Biosystems Instrumentation). Primers were designed using Primer Blast (NCBI) and de‐novo synthesized by Thermo‐Fischer. Sequences for primers are included in supplemental documentation.

For IF inflammasome visualization, cytokine/fusokine stimulation was carried out as above. Cells were then fixed (PFA), blocked (Goat Serum), and stained with anti‐ASC (find), Phalloidin (Sigma), and Dapi mounting media (Fischer). Slides were visualized on a Keyence fluorescence microscope.