Effect of reserpine on proliferation and cardiomyocyte differentiation.

<p>(A) Microscopy images of EBs at day 10 and 14 of differentiation (day 14 EBs were purified with puromycine). eGFP positive CMs containing clusters in control, DMSO (solvent control) and reserpine-treated EBs are shown (green arrow in reserpine-treated). Scale bars: 200 µm. (B) Time course of appearing spontaneously beating clusters within plated EBs at indicated time points. (C) EB proliferation was analyzed based on cross sectional areas (mm²) from phase contrast pictures captured at days 2, 4 and 10 after differentiation start (n = 3). (D) Percentage of eGFP positive CMs from control, DMSO and reserpine treated EBs at day 10 (n = 11; *** p<0.01) and day 14 (n = 7) of differentiation derived from flow cytometry. (E) Immunocytochemical staining of EBs at day 11 of differentiation for cardiac marker proteins: eGFP positive cardiomyocytes (green), Hoechst 33342 stained nuclei (blue) and cardiac αActinin (red; top panel) or cardiac TroponinC (red; bottom panel). Top row represents overlay pictures. Scale bars: 50 µm.