FlowSOM and t-SNE analyses pipelines identify a discrete population within CD14^hi monocytes coexpressing MS4A4A, CD86, SIGLEC5, and CD163 at the protein level in LKs. (A) Heat map visualization of marker expression in each of the 8 metaclusters generated by FlowSOM algorithm on concatenated monocytes data of 30 patients in bulk leukapheresis products (3000 cells per sample). The mean fluorescence intensity of each marker (MS4A4A, CD86, SIGLEC5, and CD163 columns) is displayed for each metacluster (rows), n = 30. (B) t-SNE, a nonlinear dimensionality reduction method, was used to visualize FlowSOM metaclusters. (C) t-SNE pseudocolored map representing expression of CD14 marker. Red indicates higher expression and blue indicates lower expression. (D) Percentage of metacluster 1 among monocytes in SIG^pos and SIG^neg. (E) Percentage of metacluster 1 among monocytes in RE and NR. In panels D-E, exact median values are reported. Comparisons were performed using Mann-Whitney test; ∗P < .05. FlowSOM, Flow Self-Organizing Map; t-SNE, t-distributed stochastic neighbor embedding.