Figure 1.
Identification of homozygous complete LOFATRIP variants in patients with MPD. (a) Family pedigree with allele segregation of ATRIP splice variant. Index patient (F1Pt) is marked with an arrow, and clinical phenotype and genotype are indicated in the legend. A detailed phenotypical description can be found in Materials and methods. (b) Fragment analysis and size profiles of PCR-amplified cDNA extracted from PHA blasts for F1Pt, parents (F1Fa and F1Mo), and HC. Arrows indicate the position of forward and reverse primers used for PCR amplification. Percentages represent relative quantification of the 538 bp wild type (WT) and 380 bp mutant (r.671_829del) fragment. Data are representative of five independent experiments. (c) Electropherograms of cDNA extracted from PHA blasts of F1Pt and a control. Nucleotide numbering is in accordance with ENST00000320211.1. Depicted profiles are reflective of five independent experiments. (d) Schematic illustration of biallelic ATRIP variant effect at both transcript and amino acid level. (e) Endogenous protein expression of ATRIP and complex partners in primary fibroblasts from F1Pt, F02-98 (ATR patient), and controls (HCs). β-Tubulin was used as the loading control. Western blot image is reflective of four independent experiments. (f) Peak size profile in base pairs (bp) of PCR-amplified cDNA extracted from EBV-LCLs generated from the father (F1Fa) treated with and without puromycin. Percentages represent the relative ratio of WT (835 bp) and mutant (380 bp) fragments. Data are reflective of two independent experiments. (g) Schematic overview of the effect of the biallelic ATRIP variant at the protein level and an overview of the ATR-ATRIP complex with downstream effector CHK1. (h) HEK293T cells were transiently co-transfected with 3xFLAG-tagged ATRIP (3xFLAG-wtATRIP or 3xFLAG-mutATRIP), RPA70-Myc, and ATR-V5. After immunoprecipitation with anti-FLAG or control IgG, the interaction between ATRIP (FLAG) and ATR (V5), RPA70 (Myc), and endogenous TOPBP1 was examined by western blot analysis. β-Tubulin serves as a loading control. Results are reflective of three independent experiments. Source data are available for this figure: SourceData F1. Refer to the image caption for details.

Identification of homozygous complete LOF ATRIP variants in patients with MPD. (a) Family pedigree with allele segregation of ATRIP splice variant. Index patient (F1Pt) is marked with an arrow, and clinical phenotype and genotype are indicated in the legend. A detailed phenotypical description can be found in Materials and methods. (b) Fragment analysis and size profiles of PCR-amplified cDNA extracted from PHA blasts for F1Pt, parents (F1Fa and F1Mo), and HC. Arrows indicate the position of forward and reverse primers used for PCR amplification. Percentages represent relative quantification of the 538 bp wild type (WT) and 380 bp mutant (r.671_829del) fragment. Data are representative of five independent experiments. (c) Electropherograms of cDNA extracted from PHA blasts of F1Pt and a control. Nucleotide numbering is in accordance with ENST00000320211.1. Depicted profiles are reflective of five independent experiments. (d) Schematic illustration of biallelic ATRIP variant effect at both transcript and amino acid level. (e) Endogenous protein expression of ATRIP and complex partners in primary fibroblasts from F1Pt, F02-98 (ATR patient), and controls (HCs). β-Tubulin was used as the loading control. Western blot image is reflective of four independent experiments. (f) Peak size profile in base pairs (bp) of PCR-amplified cDNA extracted from EBV-LCLs generated from the father (F1Fa) treated with and without puromycin. Percentages represent the relative ratio of WT (835 bp) and mutant (380 bp) fragments. Data are reflective of two independent experiments. (g) Schematic overview of the effect of the biallelic ATRIP variant at the protein level and an overview of the ATR-ATRIP complex with downstream effector CHK1. (h) HEK293T cells were transiently co-transfected with 3xFLAG-tagged ATRIP (3xFLAG-wtATRIP or 3xFLAG-mutATRIP), RPA70-Myc, and ATR-V5. After immunoprecipitation with anti-FLAG or control IgG, the interaction between ATRIP (FLAG) and ATR (V5), RPA70 (Myc), and endogenous TOPBP1 was examined by western blot analysis. β-Tubulin serves as a loading control. Results are reflective of three independent experiments. Source data are available for this figure: SourceData F1.

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