Inducing Apoptosis in Jurkat T Cells

Jurkat human T cell line was kindly provided by Prof. J. Smit (UMCG) and cultured in RPMI (Gibco, 21,875,034) supplemented with 10% FBS, 1 × penicillin–streptomycin, and 1 mM sodium pyruvate. Apoptosis induction for phagocytosis assay (described below) was done on the day of performing the assay as previously described [2 (link)]. Jurkat cells were collected at 1*10⁶ cells/mL in medium and treated with 1 μM staurosporine (Sigma-Aldrich, S5921) for 4 h. To verify induction of apoptosis, Jurkat cells were labelled with Annexin V/propidium iodide (Biolegend, 640,914) according to manufacturer’s instructions and analyzed on a MacsQuant (Miltenyi Biotec). After 4 h of staurosporine exposure, > 90% of cells were early apoptotic (Additional file 7: Fig. S6D).