Extended Data Fig. 6.

a-b Reconstructed 3D images of nuclei retrieved from images of WT and MLL4^P4093X MSCs (a) or primary fibroblasts from healthy donor and Kabuki patients (b). Scale bar, 5 μm. The nuclear area, volume and flattening were determined and represented as box plots indicating the median (middle line), the first and third quartiles (box), and the 10th and 90th percentile (error bars). The number of analyzed nuclei is reported in figure as n; unpaired two-tailed Student’s t-test was applied for statistical analysis (****P<0.0001).

c Measurements of nuclear shape in WT and MLL4^Q4092X MSCs expressing CRISPRa with or without crRNA targeting KMT2D promoter. The nuclear area, volume and flattening were represented as box plots indicating the median (middle line), the first and third quartiles (box), and the 10th and 90th percentile (error bars). The number of analyzed nuclei is reported in figure as n; unpaired two-tailed Student’s t-test was applied for statistical analysis (****P<0.0001).

d qRT-PCR of LMNA in WT and MLL4^Q4092X MSCs, normalized on GAPDH level. Data are means + SEM (n=3 independent experiments); unpaired two-tailed Student’s t-test was applied for statistical analysis.

e Quantifications of immunostaining for LMNA and phosphorylated LMNA/C (pLMNA) in WT and MLL4^P4093X MSCs. Box plots indicate the median (middle line), the first and third quartiles (box), and the 10th and 90th percentile (error bars) of the fluorescence intensity. The number of analyzed nuclei is reported in figure as n; unpaired two-tailed Student’s t-test was applied for statistical analysis (****P<0.0001).

f-g Representative images and quantifications of cellular area detected by Phalloidin staining in WT and MLL4^Q4092X MSCs (f) or in the same cells expressing either EGFP or EGFP-Nesprin-KASH (g). Scale bar, 20 μm. Box plots indicate the median (middle line), the first and third quartiles (box), and the 10th and 90th percentile (error bars) of the cellular area. The number of analyzed cells is reported in figure as n; unpaired two-tailed Student’s t-test was applied for statistical analysis (****P<0.0001).

h-i Representative images and quantifications of immunostaining for H4K16ac in WT and MLL4^P4093X MSCs (h) or in WT primary fibroblasts from healthy donor or Kabuki patients (i). Box plots indicate the median (middle line), the first and third quartiles (box), and the 10th and 90th percentile (error bars) of the normalized fluorescence intensity. The number of analyzed nuclei is reported in figure as n; unpaired two-tailed Student’s t-test was applied for statistical analysis (****P<0.0001).