Skip to main content
. 2021 Apr 16;296:100674. doi: 10.1016/j.jbc.2021.100674

Figure 3.

Figure 3

Effect of the PLD inhibitor, FIPI, or the Gqinhibitor YM254890 on the translocation of WT GFP-mSK1 in MCF-7L cells. MCF-7L cells overexpressing WT GFP-mSK1 were pretreated with and without FIPI (100 nM, 1 h) or YM254890 (10 μM, 30 min) before S1P (5 μM) or PMA (1 μM) or carbachol (100 μM) for 10 min. Cells were processed and mounted with DAPI to stain the DNA (blue). A, photomicrographs of cells of 40× oil magnification overexpressing WT GFP-mSK1 detected with GFP. Representative results of three independent experiments. B, the bar graphs (lower of each pair) represent the AUC of transfected WT GFP-mSK1 translocation (n = 5) and (upper of each pair) the percentage of cells containing translocated WT GFP-mSK1 (n = 3); ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001 for stimulus alone versus stimulus with either FIPI or YM254890; +p < 0.05, ++p < 0.01, +++p < 0.001, and ++++p < 0.0001 for stimulus versus control transfected WT GFP-mSK1 (two-way ANOVA with Tukey's post hoc test). AUC, area under the curve; FIPI, N-[2-[4-(2,3-dihydro-2-oxo-1H-benzimidazol-1-yl)-1-piperidinyl]ethyl]-5-fluoro-1H-indole-2-carboxamide hydrochloride; mSK1, mouse SK1; PLD, phospholipase D; PMA, phorbol 12-myristate 13-acetate; S1P, sphingosine-1-phosphate; SK1, sphingosine kinase 1.