Extended Data Fig. 5: KMT2D truncating mutations affect PcG clustering. | Nature Genetics

Extended Data Fig. 5: KMT2D truncating mutations affect PcG clustering.

From: MLL4-associated condensates counterbalance Polycomb-mediated nuclear mechanical stress in Kabuki syndrome

Extended Data Fig. 5: KMT2D truncating mutations affect PcG clustering.The alternative text for this image may have been generated using AI.

a, Western Blot analysis of EED, EZH2 and SUZ12 in WT and MLL4Q4092X MSCs; histone H3 was used as loading control. b, qRT-PCR of BMI and RING1B in WT and MLL4Q4092X 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. c, Quantifications of immunostaining for H3K27me3, BMI, and RING1B in WT and MLL4P4093X 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). d–f, Representative images and quantifications of immunostaining for H3K27me3 (d), BMI (e), and RING1B (f) in primary fibroblasts from healthy donor or Kabuki patients; 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 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). g, qRT-PCR of KMT2D in WT and MLL4Q4092X MSCs, expressing CRISPRa with or without crRNA targeting KMT2D promoter. Retrieved data were normalized on GAPDH level. Data are means + SEM (n=4 independent experiments); unpaired two-tailed Student’s t-test was applied for statistical analysis. h, i, Quantifications of immunostaining for MLL4, H3K4me1, BMI, RING1B and H3K27me3 in WT and MLL4Q4092X MSCs expressing CRISPRa with or without crRNA targeting KMT2D promoter. 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; Student’s two-tailed unpaired t-test was applied for statistical analysis (****P<0.0001).

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