Additional file 3 of Immuno-priming durvalumab with bevacizumab in HER2-negative advanced breast cancer: a pilot clinical trial

Additional file 3 : Supplementary Figure S3: Flow cytometry gating strategy for innate cell populations. Representative example of the gating strategy used for immunophenotyping of the indicated innate subpopulations. After the exclusion of dead cells, monocytes were selected by size and complexity; subtypes were identified by CD14 and CD16 staining, allowing the identification of classical monocytes (CD14+ CD16−), alternative monocytes (CD14−CD16+) and intermediate monocytes. For DC and NK, leukocytes were selected by size and complexity in the live cells, and T- and B-lymphocytes excluded by staining with lineage-specific antibodies. Dendritic cells (DC) and NK cells were selected within the CD20− and CD14− population; NK cells were identified as CD56+ cells (both CD16+ and CD16−) and DC as HLA-DR+CD16− cells. DC subtypes were further defined by CD11c+ (myeloid DC) or CD123+ (plasmacytoid DC).