Figure 150.

Gating strategy for LSEC identification in liver samples isolated from C57BL/6 mice (9 weeks-old). Single cells were discriminated by plotting FSC-A vs FSC-H and SSC-A vs SSC-H (gate 1 and 2, respectively). Then, the population of interest were gated based on its morphology (gate 3). Living cells were defined as Live/Dead NIR negative (gate 4). Kupffer cells and macrophage-related population were excluded using F4/80 (gate 5). Other leukocytes (e.g., lymphocytes, NKT cells, ILCs, and neutrophils) were excluded using CD45.2 (gate 6). LSEC are finally identified as CD31⁺CD146⁺(gate 7).