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. 2021 Apr 16;296:100674. doi: 10.1016/j.jbc.2021.100674

Figure 2.

Figure 2

Colocalization of SK1 with markers of lamellipodia and filopodia. MCF-7L cells overexpressing WT GFP-mSK1 were treated with 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, the bar graph representing the percentage of cells containing translocated WT GFP-mSK1 in lamellipodia and filopodia microdomains in the PM in response to stimulus (n = 4); ∗∗∗∗p < 0.0001 treated versus control (two-way ANOVA with Tukey's post hoc test). B, photomicrographs of cells of 40× oil magnification overexpressing WT GFP-mSK1 detected with GFP and cortactin (marker of lamellipodia), fascin (marker of filopodia), and paxillin (marker of adhesion foci) detected with respective specific antibodies. Representative results of three independent experiments. Also shown is a bar graph of the Pearson Correlation Coefficients of colocalization. ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001 for Pearson Correlation Coefficients for stimulated versus control (increase/decrease) using two-way ANOVA with Tukey's post hoc test. mSK1, mouse SK1; PM, plasma membrane; PMA, phorbol 12-myristate 13-acetate; S1P, sphingosine-1-phosphate; SK1, sphingosine kinase 1.