Fig. 5: Chromatin is detached from the NE upon Elys-KD.

a Confocal images of FISH signals (red) detected by the probe for 60D region in nuclei stained with anti-Lamin (green) and anti-LBR (violet) antibodies in Elys-KD or control S2 cells. Scale bar 1 µm. b–d Violin-plots showing distribution of radial-normalized distances between 60D (b), 22A (c), or 36C (d) FISH signals and the NE in control (blue; n = 141, or n = 151, or n = 125 for 60D, 22A and 36C probes, respectively), Elys-KD (red; n = 137, or n = 150, or n = 125 for 60D, 22A and 36C probes, respectively), Lam-KD (brown; n = 100), and simultaneous Elys-KD and Lam-KD (dark brown; n = 100) S2 cells. e, f Averaged fluorescence intensity profiles along the nuclear diameter in Elys-KD (n = 50) and control (n = 50) S2 cells immunostained with anti-histone H4 and anti-LBR antibodies (e), or in Elys-KD (n = 170) and control (n = 170) S2 cells immunostained with anti-H3K27Ac and anti-Lamin antibodies (f). To estimate P-values, two fluorescence intensity distributions (control vs Elys-KD) on the interval of radial positions from 0 (position of the NE identified by the peak of LBR or Lamin Dm0 staining) to 0.15R, were compared. This interval is delimited by the dashed lines. g Violin-plots showing the distribution of volumes of control (n = 250) and Elys-KD (n = 250) S2 nuclei manually outlined by the NL and further automatically reconstructed in IMARIS. White dots represent median values, upper and lower ends of black bars show the upper and lower quartiles, the ends of the thin line indicate the maximum and minimum values. h A schematic illustrating mechanisms of peripheral chromatin attachment to the NE. All measurements in (b–g) were performed in two biological replicates. P-values in (b–g) were estimated in an M-W U-test.