FIGURE 2.

Modulation of the S1P lyase (SPL) expression level influences cell proliferation, migration, invasion and glyceroLPL levels mostly in glucose absent conditions. (A, B and E) Confirmation of stably overexpressed or inhibited SPL in the mouse colon cancer cell line Colon 26 by measuring the (A) mRNA expression levels (N = 3) and (B and E) protein levels. The mammalian cell expression vector p3FLAG CMV‐10 was used to induce stable overexpression, and the CRISPR/Cas9 system for stable inhibition. (C, D, F and G) The cell proliferation (N = 24), migration (N = 3) and invasion (N = 3) rates evaluated in the SPL‐overexpressing (C and D) and SPL‐inhibited cell lines (F and G), in the presence or absence of glucose. (H and J) The total glycero‐LPL levels in the SPL‐overexpressing and SPL‐inhibited cell lines. (I and K) S1P levels in cell lines in which the SPL expression levels were modulated. Measurement of the total glycero‐LPL and S1P levels (N = 6) were performed in the cell lysates and adjusted to the protein concentrations (which were used later for the analysis). Differences between the two groups were assessed using the unpaired Student's t‐test, while differences among more than two groups were assessed using a one‐way ANOVA, p < .05 was considered as being indicative of statistical significance. EV, empty vector transfected control cell line; HR, homologous recombination vector only transfected control cell line; +SPL, SPL overexpressing cell line; Cr/SPL, SPL knock out cell lines by using CRIPR/Cas9 system