Fig. 6

Role of fluid shear on hESC priming, cell cycle, and apoptosis. a, b POU5F1 (a) and NANOG (b) expressions obtained from qPCR analysis. Data were normalized to GAPDH and presented as the mean ± SE in three replicates under each condition. c, d POU5F1 (c) and NANOG (d) immunostaining for hESCs under static and fluid shear conditions. Data in right columns were presented as the mean ± SE obtained from three replicates for total 60 colonies under each condition. e Representative images of nucleolus (indicated by thick white arrow in the most right column) for a typical colony. f, g DNA at different mitotic phases was visualized with Hoechst 33342 (blue) (f) to estimate the percentage of mitotic cells (g). Also plotted was the apoptotic percentage of hESCs using FACS analysis (h). The numbers of test replicates and measured colonies, (M, m), are (4, 32) for static control, (3, 36) for fluid shear, and (3, 30) for TSA treatment in g. Bar = 10 μm in e and 20 μm in f. Here Shear denotes the steady shear of 1.1 Pa for 24 h