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. 2018 Oct 10;8:376. doi: 10.3389/fonc.2018.00376

Figure 5.

Figure 5

Depletion of ABCG1 triggered accumulation of EVs and declined survival of tumor cells. (A–E) Depletion of ABCG1 increased accumulation of EV-derived lipid in LuM1 cells in 2D-culture condition (A–D) and 3D tumoroids (E). LuM1-derived EVs were labeled with fluorescent sphingolipid and added to conditioned media of ABCG1-depleted or control LuM1 cells. (A) Accumulation levels of EV-lipid. Accumulation levels per cell at 3 h post-EV addition period were shown. **P = 0.0004, n = 4 (biological replicates). (B) The rate of EV-lipid-accumulated cells. The rate of EV-lipid accumulated cells at 3 h post-EV addition period was shown. **P = 0.0003, n = 4 (biological replicates). (C) Kinetics of EV accumulation in the 2D-cultured LuM1. **P = 0.0004, n = 4 (biological replicates). (D) Representative images of cells with EV-lipid accumulation at 3 or 24 h post-EV addition periods. Red fluorescence indicates EV-lipid. DNA was stained with Hoechst33342 (blue). Scale bars, 200 μm. (E) Accumulation levels of EV-lipid in 3D tumoroids. *P < 0.05, **P < 0.01, ***P < 0.001, n = 8 (biological replicates).