Extended Data Fig. 6: Flow cytometry gating scheme for whole blood and T cell assessment in MDM with and without activation.
From: Monocyte-derived macrophages contain persistent latent HIV reservoirs
Post singlet gating samples are gated on TLR2 and side scatter to separate monocytes (TLR2+) from lymphocytes (TLR2−) (A). TLR2+ cells are then gated in monocytes subsets, classical (CD14 + CD16−), intermediate (CD14 + CD16+) and non-classical (CD14lo/-CD16+) (B). TLR2- cells are separated based on CD3 and CD159a expression (C) and then further gate on CD4 and CD8 expression (D). CD4 T cells are gated as (TLR2-CD3 + CD4 + CD8−). (E) MDM with and without activation for 12 days with PMA were removed from the plate with TrypLE and stained with Live/Dead Near IR and with or without CD3 for 30 minutes at 4C. Cells were permeabilized using Biolegend PermFast and stained with CD68 or matched IgG control. Cells were run immediately on a BD LSRFortessa. Cells were first gated to remove debris, then to remove doublets, and finally to remove dead cells.
