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. 2021 Aug 7;11(8):1169. doi: 10.3390/biom11081169

Figure 1.

Figure 1

Gating strategy for characterizing human monocyte subsets. White blood cells were first gated on a time/side scatter (SS) density plot, and then gated on the Zombie UV negative cell population for detection of living cells. Afterwards, living cells were gated for singlets on a forward scatter (FS)/Trigger Pulse Width density plot. Monocytes were recognized on the HLA-DR gating. Then, monocytes were gated on the CD14+/CD16+ population for identification of CMs (CD14++CD16−), IMs (CD14++CD16+), and NCMs (CD14+CD16+). Expression of IL-1 beta, CCR2, and CX3CR1 was measured in all monocyte subsets. SSC, side scatter; FSC, forward scatter; HLA-DR, human leukocyte antigen-DR isotype; CM, classical monocytes; IM, intermediate monocytes; NCM, non-classical monocytes; IL-1 beta, interleukin 1 beta; CCR2, C-C chemokine receptor type 2; CX3CR1, CX3C chemokine receptor 1.