(A) TINF2 mutations and clinical features of affected individuals in four different families. Telomere length percentile is based on Flow-FISH data (see below Figure 5—figure supplement 1A). (B, C) …
Pedigrees of two families with heterozygous c.604G > C mutations listed in Figure 1A. Probands are highlighted by arrows. Filled symbols indicate patients with confirmed TINF2 mutations and clinical …
Table showing the somatic mutations identified in the tumors of families with TINF2 mutations.
(A) Transcript analysis in peripheral blood lymphocytes (with and without cycloheximide treatment, CHX) from patients with the c.604G > C TINF2 mutation (F1:III-3 and F1:II-4; see Figure 1A) and a …
(A) Co-immunoprecipitation of myc-tagged TRF1 (left panel), TRF2 (middle panel) and TPP1 (right panel) from 293T cells co-transfected with HA-tagged wt TIN2, S186fs, L170fs, E202fs, or the empty …
(A) Immunoblot for TIN2 and γtubulin in control cells and the indicated clones with targeted TINF2 alleles. (B) Quantification of the immunoblot shown in A. Unpaired t-test was used to determine …
Transcript analysis in control cells and cells with c.604G > C TINF2 mutations. Wild-type and alternative (604G > C I, 604G > C II, alt. splice exons 3–6) transcripts are indicated. Alt. splice …
(A) Schematic of the TINF2 locus showing landmarks relevant to CRISPR/Cas9-mediated knock-in of TINF2 mutations. The single guide RNA (sgRNA) target regions and the primers used for genotyping (blue …
(A) Schematic of the TINF2 locus showing landmarks relevant to CRISPR/Cas9-mediated targeting of exon1. The single guide RNA (sgRNA) target region and the primers used for genotyping (blue arrows) …
(A–C) Reference sequence with sgRNA sequence and PAM and the edited sequences (highlighted) of the obtained cell lines for c.557del (A), c.604G > C (B) and TIN2+/- cells (C) and control cell lines.
(A) Growth curves of RPE1 control cells and cells with heterozygous c.557del or c.604G > C, homozygous c.604G > C mutations, or TIN2+/- cells. (B) Telomeric ChIP to determine the specificity of Abs …
(A) Quantification of telomeres containing 53BP1 (≥50 nuclei per cell line, with individual cell lines from Figure 4D shown). (B) Quantification of telomere fusions (≥20 spreads per cell line, with …
(A) Telomeric Southern blot of MboI/AluI-digested genomic DNA from immortalized and primary patient cells (lymphocytes), normal lung fibroblasts (MRC5), and HeLa1.3 cells. Median telomere length …
(A) Median telomere length (MTL) in individuals with TINF2 truncations as measured by Flow FISH. (B) Schematic of the TINF2 locus showing landmarks relevant to CRISPR/Cas9 mediated targeting of the …
Quantification of telomeric sister chromatid exchanges (T-SCE) in RPE1 control cells, cells with heterozygous c.604G > C, or TIN2+/- cells (≥15 spreads per cell line, and two cell lines per …
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Cell line (H. sapiens) | 293T | ATCC | ||
Cell line (H. sapiens) | hTERT-RPE1 p53-/- Rb-/- | Yang et al., 2017 | ||
Cell line (H. sapiens) | hTERT-RPE1 p53-/- Rb-/- +/604G > C.1 clone 1–3 m | This paper | Heterozygous forTINF2 c.604G > C | |
Cell line (H. sapiens) | hTERT-RPE1 p53-/- Rb-/- +/604G > C.2 clone 2–23 m | This paper | Heterozygous forTINF2 c.604G > C | |
Cell line (H. sapiens) | hTERT-RPE1 p53-/- Rb-/- +/604G > C.3 clone 1–21 m | This paper | Heterozygous forTINF2 c.604G > C | |
Cell line (H. sapiens) | hTERT-RPE1 p53-/- Rb-/- ctrl4 clone 1–4 c | This paper | Control cell line | |
Cell line (H. sapiens) | hTERT-RPE1 p53-/- Rb-/- ctrl5 clone 1–13 c | This paper | Control cell line | |
Cell line (H. sapiens) | hTERT-RPE1 p53-/- Rb-/- ctrl6 clone 5–8 c | This paper | Control cell line | |
Cell line (H. sapiens) | hTERT-RPE1 p53-/- Rb-/- +/557del.1 clone 1–9 m | This paper | Heterozygous forTINF2 c.557del | |
Cell line (H. sapiens) | hTERT-RPE1 p53-/- Rb-/- +/557del.2 clone 1–14 m | This paper | Heterozygous forTINF2 c.557del | |
Cell line (H. sapiens) | hTERT-RPE1 p53-/- Rb-/- +/557del.3 clone 2–17 m | This paper | Heterozygous forTINF2 c.557del | |
Cell line (H. sapiens) | hTERT-RPE1 p53-/- Rb-/- ctrl1 clone 2–7 c | This paper | Control cell line | |
Cell line (H. sapiens) | hTERT-RPE1 p53-/- Rb-/- ctrl2 clone 2–8 c | This paper | Control cell line | |
Cell line (H. sapiens) | hTERT-RPE1 p53-/- Rb-/- ctrl3 clone 1–2 c | This paper | Control cell line | |
Cell line (H. sapiens) | hTERT-RPE1 p53-/- Rb-/- +/-.1 clone 1-1het | This paper | Heterozygous for TINF2 | |
Cell line (H. sapiens) | hTERT-RPE1 p53-/- Rb-/- +/-.2 clone 1-3het | This paper | Heterozygous for TINF2 | |
Cell line (H. sapiens) | hTERT-RPE1 p53-/- Rb-/- +/-.3 clone 2-4het | This paper | Heterozygous for TINF2 | |
Cell line (H. sapiens) | hTERT-RPE1 p53-/- Rb-/- +/+ ctrl7 clone 2–9 c | This paper | Control cell line | |
Cell line (H. sapiens) | hTERT-RPE1 p53-/- Rb-/- +/+ ctrl8 clone 3–2 c | This paper | Control cell line | |
Cell line (H. sapiens) | hTERT-RPE1 p53-/- Rb-/- +/+ ctrl9 clone 1–4 c | This paper | Control cell line | |
Cell line (H. sapiens) | WIBR3 hESC | Lengner et al., 2010 | NIH stem cell registry number: 0079 | Wild-type |
Cell line (H. sapiens) | WIBR3 hESC 1B1 | This paper | +/+ TINF2 Control cell line | |
Cell line (H. sapiens) | WIBR3 hESC 3B4 | This paper | +/+ TINF2 Control cell line | |
Cell line (H. sapiens) | WIBR3 hESC 3C2 | This paper | +/+ TINF2 Control cell line | |
Cell line (H. sapiens) | WIBR3 hESC 5B1 | This paper | +/+ TINF2 Control cell line | |
Cell line (H. sapiens) | WIBR3 hESC 3B3 | This paper | +/- TINF2 | |
Cell line (H. sapiens) | WIBR3 hESC 4B4 | This paper | +/- TINF2 | |
Cell line (H. sapiens) | WIBR3 hESC 5A4 | This paper | +/- TINF2 | |
Cell line (H. sapiens) | WIBR3 hESC 5B3 | This paper | +/- TINF2 | |
Anti-hTIN2 | Ye and de Lange, 2004 | #864 | ||
Anti-γTubulin | Sigma | GTU88 | ||
Anti-Myc | Cell signaling | 9B11 | ||
Anti-HA | Abcam | Ab9110 | ||
Anti-53BP1 | Abcam | ab175933 | ||
Cy3-OO- (TTAGGG)3 | PNA bio | Telomere probe | ||
FITC-OO-(CCCTAA)3 | PNA bio | Telomere probe | ||
Alexa Fluor 647-OO-(TTAGGG)3 | PNA bio | Telomere probe | ||
CENPB-AF488 | PNA bio | F3004 | Centromere probe |
Uncropped images of immunoblots and telomere blots shown in the main figures.
Uncropped images of immunoblots and telomere blots shown in the main figures.
Uncropped images of immunoblots and telomere blots shown in the main figures.
p-Values and summary statistics for key data.