Extended Data Fig. 8: A patient-specific nonsense mutation in Adnp impairs the interaction with HP1 but not with DNA.

a, Scheme depicting the wild-type and mutant Adnp alleles, which code for Tyr (blue) and a patient-specific premature termination codon (red) at amino acid position 718, respectively. Full-length and truncated protein products are shown on the right. Arrow indicates transcription start site. Boxes represent exons. Numbers denote amino acids. b, N-terminally Flag-AviTag-tagged ADNP^PTC718 was streptavidin-purified from cells with and without aminoglycoside treatment (gentamycin or paromomycin) and subjected to LC–MS/MS analysis. ADNP^PTC718-expressing cells were treated with 2 mg ml⁻¹ gentamycin (2.9 mM) or paromomycin (3.2 mM) for 24 h. The table depicts total spectral counts, unique peptides and percentage sequence coverage (derived from Scaffold) for all ChAHP components from the different treatments. c, qRT–PCR measurement of Bmp1 and Igfbp4 mRNA levels in ES cells expressing full-length Adnp (Adnp^+/+) or C-terminally truncated Adnp that interacts with CHD4 but not with HP1 (Adnp^{PTC718/PTC718}). n = 3 biological replicates (that is, three independent RNA isolations). P values were calculated using two-tailed unpaired unequal variances t-tests. Centre value denotes the mean; error bars denote s.d. d, ChIP–qPCR enrichments for transiently transfected Flag-AviTag-tagged wild-type ADNP and ADNP^PTC718 constructs on two ADNP targets, normalized to an intergenic control. Black lines indicate means. e, C-terminally Flag-AviTag-tagged ADNP^PTC718 was streptavidin-purified from cells with or without gentamycin treatment (2.9 mM) and subjected to LC–MS/MS analysis. Bold letters indicate unique peptides further quantified by parallel reaction monitoring (PRM). C-terminal peptides encoded downstream of PTC718 are shown in colour. Dashed box denotes the HP1 interaction motif. f, Summed fragment intensities of five C-terminal ADNP peptides that are encoded downstream of PTC718 are shown on the left. Background proteins shown on the right serve as loading controls. Intensities were measured by PRM. Total spectrum counts were derived from Scaffold. n = 3 biological replicates.