Chunk #43 — EXPERIMENTAL PROCEDURES — Flow Cytometry

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IRE1α induces thioredoxin-interacting protein to activate the NLRP3 inflammasome and promote programmed cell death under irremediable ER stress.

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For assaying apoptosis by Annexin V staining, cells were plated two days prior to FACS in 6-well plates. The day before flow cytometry, MEFs were induced with either 5μg/ml Tm or 1μM Tg. The next day, cells were trypsinized and washed in PBS and resuspended in Annexin V binding buffer with Annexin-V FITC (#K101-100, Biovision.). Flow cytometry was performed on a Becton Dickinson LSRII flow cytometer.