Supplementary Figure 8: Defective nuclear lamina architecture and impaired nuclear mechanics upon loss of MOF and lamin A/C acetylation.

a) Representative electron tomographic slices showing preserved nuclear structure in Mof control and WT lamin A MEFs. N; nucleus. b) Quantification of electron tomographic micronuclei heterochromatin density in Mof KO and K311R lamin A MEFs. Data shown are the mean ± s.e.m. n=15 nuclear structures counted. c) Quantification of electron tomographic submembrane (120 nm) heterochromatin density in the 1-μm-vicinity of fuzzy nuclear lamina sites; Two-tailed nonparametric Mann–Whitney test. The data shown are the mean ± s.e.m. n numbers are stated in the panel and represent single cells counted. d) Quantification of electron tomographic global nuclear heterochromatin density measured; Two-tailed nonparametric Mann–Whitney test. Data shown are the mean ± s.e.m. n numbers are stated in the panel and represent single cells counted. e) Relative Emerald-lamin A FRAP over time in Lmna KO MEFs overexpressing WT or K311R lamin A. Two-tailed nonparametric Mann–Whitney test for n=45 WT and n=50 K311R cells. f) Immunoblotting on whole cell extracts of MEFs upon Mof or Kansl2 KO (4 days). Uncropped blots in Supplementary Fig. 9. g) Nuclear and cyto-indentations are performed in triplicate for each cell (left image) and for each isolated nucleus (right image). Pink crosses indicate sites of nanoindentation. Scale bar; 20μm. h) Schematics of the nanoindentation experiment performed and representative force distance curve used to calculate the effective Young’s Modulus. Fit of the indentation curve used to obtain the Young’s Modulus shown in black. i) Cytoindentation quantification on intact control and Mof KO (4 days) MEFs; Two-tailed nonparametric Mann–Whitney test. Data shown are the mean ± s.e.m. n numbers are stated in the panel and represent single cells counted. j) Nanoindentation nuclei stiffness quantification in intact control and Mof KO (4 days) MEFs; Two-tailed nonparametric Mann–Whitney test. Data shown are the mean ± s.e.m. n numbers are stated in the panel and represent single cells counted. k) Nanoindentation stiffness quantification on freshly isolated control and Mof KO (4 days) MEFs nuclei; Two-tailed nonparametric Mann–Whitney test. Data shown are the mean ± s.e.m. n numbers are stated in the panel and represent single nuclei counted.