Extended Data Fig. 1: Mechano-osmotic changes of the nucleus accompany exit from pluripotency.
From: Mechano-osmotic signals control chromatin state and fate transitions in pluripotent stem cells
(a-c) Representative top views (x-y), 3D reconstructions and cross sections (z), and quantification of nuclear height from of Sox2-GFP-tagged hiPSCs undergoing ectodermal (a), mesodermal (b), or endodermal (c) differentiation for the indicated time points (scale bars 15 µm; n = 3 independent experiments with 348, 496, 841, 765 (mesoderm 0, 8, 24, 48 h, respectively); 435, 661, 800, 835 (ectoderm 0, 8, 24, 48 h, respectively) total cells/condition; ANOVA/ Dunnett’s). (d) Quantification of lineage marker intensity from immunofluorescence stainings in hiPSCs undergoing ectodermal (left), endodermal (middle), or mesodermal (right) differentiation for the indicated time points (scale bars 15 µm; n = 3 independent experiments with 5093, 1257, 3913, 8290, 11345 (ectoderm SOX1); 1257, 3255, 5751, 8290, 11309 (ectoderm OCT4); 1362, 3967, 4680, 6542, 8450 (Endoderm OCT4); 1362, 3967, 4680, 6542, 6410 (endoderm SOX17); 841, 3089, 4897, 5368, 6189 (mesoderm Brachyury); 842, 3089, 4897, 5368, 6186 (mesoderm OCT4) total cells/0, 24, 48, 72, 96 h conditions, respectively; mean±SD). (e) Representative snapshots from live imaging data and quantification of nuclear volume from LaminB1-RFP hiPSCs treated with Calyculin A for 15 min. Note compaction of colony and nuclear deformation at colony edges (scale bars 50 µm; n = 4 independent experiments with 546 (DMSO) and 682 (Calyculin A) total nuclei/condition; Student’s t-test). (f) Representative images and quantification of p-p38 from hiPSCs treated with Calyculin A for 15 min. Note increased p38 activation at colony edges (scale bars 100 µm; n = 3 independent experiments with 14 (DMSO) and 17 (Calyculin A) total colonies/condition; Student’s t-test). Source numerical data are available in source data.
