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Figure 6
FTY720 induces apoptosis and suppresses the distant spread of human TNBC cells with high P-S1P1 T236 Levels. (a) Western blot analysis of P-S1P1 T236, S1P1, P-AKT S473, AKT, active CASPASE 3 (CASP3), and ACTIN levels in MI, NeoT, MIII, and MIV cells before and after FTY720 treatment (2 μM). (b) Brightfield images of MI, NeoT, MIII, and MIV cells at 48 h post-treatment with DMSO or FTY720 (n = 3 per group). The scale bar represents 1 µM. (c) The proportion of viable cells in response to a dose gradient of FTY720 shows a significantly reduced viability of MIII and MIV though not MI nor NeoT cells (n = 3 per group). (d) Overlay of brightfield and fluorescence images of zebrafish embryos injected with RFP+ MIII and MIV cells treated with DMSO or FTY720 (2 μM) at 3 dpt (n = 3 per group). The scale bar represents 200 µm. (e) Quantification of fluorescent intensity of tumor cells described in (d) Shows FTY720′s ability to inhibit the distant spread of TNBC cells (n = 3 per group). Data indicate mean ± SEM.