Fig. 4

T1-LAD, T2-LAD and nonLAD regions are distinctly radially positioned within the nucleus. A, B (Left) T1-LAD, T2-LAD, and nonLAD FISH probe locations in LB1 tracks in A ESCs and B cardiac myocytes. Probe indicated by blue box. Each probe in A and B assigned a unique identifier per cell type (E1-E3 and CM1-CM3; see also Additional file 2: Fig. S9A, F). (Right) Representative images of FISH foci (individual slices from Z-stack) from each tested probe region with 3D renderings of all slices to the right for ESCs A and fluorescence-activated cell sorted cardiac myocytes B. Box in the left image indicates the approximate area of the high magnification image in the middle. C Quantification (see “Methods”) of the distance of each FISH focus to the center of the nuclear lamina for the probes (noted by unique identifier) shown in A and B (see Additional file 2: Fig. S9A, B, F-H) relative to the middle of the nuclear lamina (LB1). Gray box indicates the depth of H3K9me2-marked heterochromatin at the nuclear periphery (see “Methods” and Additional file 2: Fig. S9D). D Cumulative frequency distribution of all probes imaged in study. Data in the light purple box on the left highlighted on the right. Error bars represent 1 SEM. Statistical significance of the differences in localization between T1-, T2-, and nonLAD foci in C calculated by a Kruskal-Wallis test with post hoc Dunn test for multiple comparisons. Statistical significance of differences in distribution between T1- and T2-LAD foci in D was calculated by a Kolmorgov-Smirnov test. *p<0.05, ****p<0.0001. Scale bars = 1μm