LPA_1/3 inhibition of human MSCs reduced actin polymerization and increased cell-cycle quiescence.

<p><b>A, B.</b> Phenotypic characteristics of human MSCs treated with Ki16425 or vehicle alone for 48 h after plating. Shown are phase-contrast images (panel <b>A</b>) and fluorescent images in which filamentous actin (F-actin) was visualized with green phalloidin-FITC staining and nuclei were stained with red propidium iodide (panel <b>B</b>). Scale bars, 200 µm. <b>C.</b> Western blotting analysis to evaluate the phosphorylation and activation status of focal adhesion kinase (FAK). <b>D.</b> Cell-cycle analysis. Human MSCs treated with Ki16425 or vehicle alone for 72 h were fixed and then stained for DNA and RNA with 7-AAD and pyronin Y, respectively. Their cell-cycle status was assessed based on their DNA and RNA content by flow cytometry. A representative of three experiments is shown on the left side, and the bar graph summarizes the results of the G₀ proportion on the right side. The data are presented as the means ± standard error (<i>n</i> = 3). <b>E.</b> Western blotting analysis of signaling molecules associated with the Akt pathway. For panels <b>C</b> and <b>E</b>, human MSCs were cultured in the presence or absence of Ki16425 for the indicated times prior to cell lysis.