Journal: Scientific Reports

Article Title: Hypoxic Stress Decreases c-Myc Protein Stability in Cardiac Progenitor Cells Inducing Quiescence and Compromising Their Proliferative and Vasculogenic Potential

doi: 10.1038/s41598-017-09813-x

Figure Lengend Snippet: Inhibition of c-Myc decreases cardiac progenitor cell proliferation and induces quiescence. ( A ) Representative histogram showing CPC proliferation analysis by CellTrace dye dilution 72 hours after treatment with the c-Myc inhibitor 10058-F4 (dotted line) and DMSO control (solid line, light gray) compared to baseline (solid line, dark gray). Graph indicates the CellTrace mean fluorescence intensity showing less dye dilution in samples treated with the c-Myc inhibitor relative to DMSO control, an indication of delayed growth (n = 4, *p < 0.03). ( B ) Quantification of Ki67 expression 72 hours after treatment of CPCs with c-Myc inhibitor and DMSO control by fluorescence activated cell sorting. Graph indicates the average of Ki67 mean fluorescence intensity (MFI) in control (black bars) and c-Myc inhibitor-treated CPCs (gray bars) (n = 4, *p < 0.03). ( C ) Cell cycle analysis of CPCs after c-Myc inhibition. Dot Plots show representative images of cells stained with 7-AAD and Ki67 and analyzed 72 hours later. Graph indicates the average percentage of cells in each phase of the cell cycle in DMSO control (black bars) and c-Myc inhibitor treated CPCs (gray bars) (n = 4, *p < 0.05).

Article Snippet: The effect of c-Myc inhibition on CPC growth was determined by staining cells with 1 μM of CellTrace Far Red Cell Proliferation Dye (Thermo Fisher Scientific) followed by treatment with 40 μM the c-Myc inhibitor 10058-F4 (Sigma-Aldrich, #F3680) or vehicle control (DMSO).

Techniques: Inhibition, Control, Fluorescence, Expressing, FACS, Cell Cycle Assay, Staining