Fig 2: CSNK2B variants identified in IDCS patients(A) Upper panel: genomic structure of human CSNK2B. The seven exons of CSNK2B are displayed by boxes, which are drawn to scale (1 kb = 1 cm). The filled boxes depict the open reading frame, and the open boxes show the UTRs. The introns are drawn as connecting lines of arbitrary length. Vertical lines on filled boxes indicate the positions of the variants. Lower panel: 215-amino-acid-long CK2ß protein composed of five domains along with identified variants are shown. The protein structure is constructed according to the indicated scale bar. Figure is recreated from Bibby and Litchfield (2005).12(B) Cross-species alignment showing conservation of Asp32 of CK2ß. Note that Asp32 is conserved in all species. Asterisk (*) is used for the conserved residues, colon (:) for conservative changes, and dot (.) for semiconservative changes.(C) Structural overview of the CK2a2ß2 holoenzyme along with a modeled substrate peptide and the ATP analogue AMPPNP. Dotted circle shows the wild-type amino acid Asp32 and substrate peptide in its close proximity.(D) Zoomed picture focuses on the critical neighborhood around Asp32 of CK2ß illustrating the IDCS-associated variants Asp32Asn and Asp32His. Two sulfate ions visible in the human CK2a structure PDB: 2PVR26 were drawn in ball-and-sticks representation; as outlined in the Material and methods section, these sulfate ions served as an orientation to model the p+1 and p+3 side chains of the substrate peptide DDSDDD (blue carbon atoms) into the active site.

Fig 3: PTEN loss of function in GXP GBMs.(a) Heatmap of the PI3K/AKT signalling pathway genes generated from RNA-seq analysis highlighting downregulated expression level of PTEN and upregulated expression level of CSNK2B (CK2ß) in 6 independent GXP GBMs compared with GXP NS. (b) Immunoblot analysis of PIK3CA, PIK3CB (indicated by red arrow), pan-AKT, p-AKT (S473), p-AKT (T308) (indicated by red arrow) and CCND1 in 7 independent GXP GBMs compared with WT or GXP NS, WT adult forebrain, GL261 (p53-mutated), U87MG (PTEN-null) or T98G (p53/PTEN mutated). MW; molecular weight. (c) Co-IF staining of PTEN and PCNA on primary GXP GBM5 paraffin-embedded sections, tumour cells (top panel) compared with adjacent normal tissue (middle panel) and WT adult brain tissue (bottom panel) are shown. PCNA and DAPI staining of nuclei are shown. White arrows in PCNA+PTEN merged images denote the regions enlarged in the far-right panels. Scale bars, 50 µm. (d) Immunoblot analysis of PTEN, p-PTEN (S380/T382/T383) and CK2ß in seven independent GXP GBMs compared with WT or GXP NS, WT adult forebrain, GL261, U87MG or T98G. See also Supplementary Fig. 2.

Fig 4: Clinical presentation of patients with CSNK2B variantsFront and side views of subjects 1–3 diagnosed with IDCS show strikingly similar facial gestalt, whereas subjects 4 and 5 diagnosed with POBINDS show vivid difference of facial gestalt, and no obvious digital or limb anomaly was recorded as in IDCS. Digital anomalies of IDCS patients are shown in two panels on right side. For subject 1, arrows show brachydactyly 2–5 and syndactyly 2–3 of the digits of the feet. Notably, preoperative radiographs of the hands and teeth are shown for subject 1.