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. 2020 Mar 4;6(10):eaaw7853. doi: 10.1126/sciadv.aaw7853

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Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).

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Fig. 4 — (A and B) The detection of nuclei within MBs enables the construction of a Voronoi diagram (A) that allows the identification of concentric cell layers (B) within the MBs. (C and D) Representative image (C) and quantitative analysis (D) (error bars represent the SD) of RUNX-2 staining within the cell layers of the MB (N_chips = 3 and n_MBs = 458). N.S., nonsignificant. (E to H) Quantitative analysis (E) and representative images (F to H) of N-cadherin staining after methanol/acetone (F) (N_chips = 3 and n_MBs = 405), after PFA/Triton X-100 fixation and permeabilization (G) (N_chips = 3 and n_MBs = 649), and F-actin staining with phalloidin (H) (N_chips = 3 and n_MBs = 421). Scale bars, 20 μm. The images were acquired using a wide-field microscope. ***P < 0.001. (I) Schematized representation of the structural organization of MBs.