Fig. 3

Characterization of spatial self-organization in the hiPSC colony with and without the ring-barrier. A Representative images in the XY plane showing pMLC and F-actin within the hiPSC colony in cultures with (a–c) the ring-barrier at day 0, and with (d–f) or without (g–i) the ring-barrier at day 2. The hiPSC colonies were compared after further exposure to HA in culture conditions without the ring barrier (j–l) at day 2. Images show the whole colony. Scale bar, 500 μm. Panels a1–l1 are the tomograms sectioned at the XZ plane (yellow dashed lines) in panels a-l. Red dashed lines indicate location of the ring culture system. Scale bar, 50 μm. B Quantification of staining intensity of pMLC and F-actin signals across the hiPSC colony with or without ring-barrier. Data are average ± S.D. of 5 cross-sections from the colonies. Significance was determined by one-way ANOVA with Tukey's test (**P < 0.01, *P < 0.05). All experiments were repeated independently at least three times with similar results