Journal: Science Advances

Article Title: Naked mole rat TRF1 safeguards glycolytic capacity and telomere replication under low oxygen

doi: 10.1126/sciadv.abe0174

Figure Lengend Snippet: ( A and B ) Representative images (A) and quantification (B and C) of colocalization between flag-TRF1 foci (green) and telomeric repeats foci (purple) under atmospheric O 2 and 3% O 2 for MEFs with TRF1 F/F + CRE + ectopic TRF1 variants (B) and for MEFs with p53 −/− CRISPR-Cas9 WT and “GLAD” ( C ) . ( D ) Representative image and ( E and F ) quantification of ChIP analyses for TRF1 binding to telomeric repeats (E) or to SINE B elements (F) in MEFs with SV40-LT TRF1 F/F + CRE + TRF1 rescue based on NMR or mouse TRF1 orthologs or TRF1 variants. Immunoprecipitation with flag antibody; bead controls were spotted onto slot blots and hybridized with a telomeric probe. Quantification of four independent experiments (in duplicate for each) that were performed for NMR and mouse TRF1 samples. Two independent experiments (in duplicate) were performed for mutGLAD and for telomeric repeats, and one experiment (in duplicate) was performed for other samples. Statistical analyses: Student’s t test, n ≥ 30 samples, * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, and **** P ≤ 0.0001; n.s. not significant.

Article Snippet: Human MRC5 fibroblasts (ATCC; #CCL-171) were immortalized by pLenti CMV/TO SV40 small + large T (Addgene, #22298) (SV40-tT MRC5).

Techniques: CRISPR, Binding Assay, Immunoprecipitation

Journal: Science Advances

Article Title: Naked mole rat TRF1 safeguards glycolytic capacity and telomere replication under low oxygen

doi: 10.1126/sciadv.abe0174

Figure Lengend Snippet: ( A and D ) Quantification of TIF of cells under 3% O 2 and after aphidicolin treatment revealed by γH2A.X staining (A) or without drug treatment (D). ( B and C ) Representative images (B) and quantification (C) of the MTS rate per chromosome spreads. Cells for (A) to (D) experiences: MEFs with SV40-LT TRF1 F/F + CRE + TRF1 rescue based on NMR or mouse TRF1 orthologs or TRF1 variants. ( E ) Quantification of TIFs, without the drug and with aphidicolin (+ APH) treatment revealed by γH2A.X staining at atmospheric oxygen and under 3% oxygen in MEFs with p53 −/− CRISPR-Cas9 WT and GLAD. Statistical analyses: Student, Wilcoxon-Mann-Whitney, and ANOVA tests, n ≥ 30 samples, * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, and **** P ≤ 0.0001.

Article Snippet: Human MRC5 fibroblasts (ATCC; #CCL-171) were immortalized by pLenti CMV/TO SV40 small + large T (Addgene, #22298) (SV40-tT MRC5).

Techniques: Staining, CRISPR, MANN-WHITNEY

Journal: Science Advances

Article Title: Naked mole rat TRF1 safeguards glycolytic capacity and telomere replication under low oxygen

doi: 10.1126/sciadv.abe0174

Figure Lengend Snippet: ( A to D ) Representative images (A) and quantification (B and D) of colocalization between flag-TRF1 foci (green) and myc-TIN2 (purple) (B); quantification of total number of flag-TRF1 foci in nucleus without and with ectopic myc-TIN2 expression (C) and of total number of myc-TIN2 foci in nucleus (D) in MEFs with SV40-LT TRF1 F/F following KO of endogenous TRF1 by CRE and rescue by ectopic NMR or mouse TRF1 orthologs under atmospheric oxygen or at 3% oxygen. ( E ) Telomere length distribution averages are represented by the shape of each beanplot, with small vertical lines that represent the mean of telomere length for each metaphase spread. Statistics: Student and ANOVA tests, n ≥ 30 samples, * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, and **** P ≤ 0.0001. AU, arbitrary units.

Article Snippet: Human MRC5 fibroblasts (ATCC; #CCL-171) were immortalized by pLenti CMV/TO SV40 small + large T (Addgene, #22298) (SV40-tT MRC5).

Techniques: Expressing