Anti cd3 eflour450

Uninfected CD4 T cells were isolated as above and treated or not with HDACi at the indicated doses. Meanwhile NK cells were isolated as above. NK and CD4 T cells were cultured at a 1:1, 1:0.2, or 1:0.01 ratio for 5 hours at 37°C in the presence of an anti-CD107a PE-Cy7 antibody (Biolegend). Experiments using K562 cells were performed at a 1:1 ratio for 5 hours as with CD4 T cells. For both CD4 T and K562 experiments, cells were then washed and stained with a panel of: LIVE/DEAD fixable near-IR dead cell stain kit (Life Technologies), anti-CD3 eflour450 (eBioscience), anti-CD56 APC (Miltenyi), anti-CD16 FITC (Biolegend), anti-CD14 APC-Cy7 (Biolegend), anti-CD19 APC Cy7 (Biolegend). NK cells were gated on APC Cy7 negative cells (live, CD14-, CD19-), CD3 negative, CD56 positive cells.

PBMC were cultured for 24h in the absence or presence of 100nM panobinostat, 20nM panobinostat, or 10nM romidepsin. After culture, wells were stained with a panel of: LIVE/DEAD fixable near-IR dead cell stain kit (Life Technologies), anti-CD3 eflour450 (eBioscience), anti-CD16 APC (Cambridge Bioscience), anti-CD56 FITC (Cambridge Bioscience), anti-CD14 APC-Cy7 (Biolegend), anti-CD19 APC Cy7 (Biolegend), NKG2D PECy7 (Cambridge Bioscience), and CD335 (NKp46) PE (Miltenyi Biotec). NK cells were gated on APC Cy7 negative cells (live, CD14-, CD19), CD3 negative, CD56 positive cells.