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<t>ESCO2</t> deficiency results in telomere abnormalities. ( A ) HeLa1.2.11 cells transfected with indicated siRNAs were analyzed by Q-FISH using a FITC-labeled telomere PNA probe. DAPI was used to stain the nuclei. The histograms show the distribution of relative telomere length presented as fluorescence intensity (TFU, telomere fluorescence unit); the red lines mark the mean telomere signal intensity. n indicates the total number of telomere signals detected. Error bars indicate standard errors. *** p < 0.001. ( B ) Examples of telomere abnormalities from ( A ) observed in a telomere FISH assay (upper panel). The incidence of telomere abnormalities in cells lacking ESCO1 or ESCO2 is shown in the bottom panel. ( C ) Sankey diagram showing KEGG pathway analysis of high-confidence proteins associated with ESCO2. Pathway enrichment was performed based on high-confidence ESCO2-interacting proteins, and the results are visualized as a Sankey diagram to illustrate the functional distribution across different KEGG pathways. ( D ) Chord diagram showing KEGG pathway analysis of DNA replication- and repair-related pathways and their associated proteins. Diagram visualizes the relationship between key proteins and their corresponding pathways involved in DNA replication and repair. ( E ) Selected lists of ESCO2-associated proteins analyzed by mass spectrometry. ( F ) Western blot performed to determine ESCO2 and <t>BLM</t> siRNA-knockdown efficiency in HeLa1.2.11 cells. ( G ) Representative images of metaphase telomere FISH in cells from ( G ) (upper panel). The incidence of telomere abnormalities is shown in the bottom panel.
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ESCO2 deficiency results in telomere abnormalities. ( A ) HeLa1.2.11 cells transfected with indicated siRNAs were analyzed by Q-FISH using a FITC-labeled telomere PNA probe. DAPI was used to stain the nuclei. The histograms show the distribution of relative telomere length presented as fluorescence intensity (TFU, telomere fluorescence unit); the red lines mark the mean telomere signal intensity. n indicates the total number of telomere signals detected. Error bars indicate standard errors. *** p < 0.001. ( B ) Examples of telomere abnormalities from ( A ) observed in a telomere FISH assay (upper panel). The incidence of telomere abnormalities in cells lacking ESCO1 or ESCO2 is shown in the bottom panel. ( C ) Sankey diagram showing KEGG pathway analysis of high-confidence proteins associated with ESCO2. Pathway enrichment was performed based on high-confidence ESCO2-interacting proteins, and the results are visualized as a Sankey diagram to illustrate the functional distribution across different KEGG pathways. ( D ) Chord diagram showing KEGG pathway analysis of DNA replication- and repair-related pathways and their associated proteins. Diagram visualizes the relationship between key proteins and their corresponding pathways involved in DNA replication and repair. ( E ) Selected lists of ESCO2-associated proteins analyzed by mass spectrometry. ( F ) Western blot performed to determine ESCO2 and BLM siRNA-knockdown efficiency in HeLa1.2.11 cells. ( G ) Representative images of metaphase telomere FISH in cells from ( G ) (upper panel). The incidence of telomere abnormalities is shown in the bottom panel.

Journal: International Journal of Molecular Sciences

Article Title: ESCO2 Interacts with TRF1/2 and Facilitates Telomere Maintenance

doi: 10.3390/ijms27062635

Figure Lengend Snippet: ESCO2 deficiency results in telomere abnormalities. ( A ) HeLa1.2.11 cells transfected with indicated siRNAs were analyzed by Q-FISH using a FITC-labeled telomere PNA probe. DAPI was used to stain the nuclei. The histograms show the distribution of relative telomere length presented as fluorescence intensity (TFU, telomere fluorescence unit); the red lines mark the mean telomere signal intensity. n indicates the total number of telomere signals detected. Error bars indicate standard errors. *** p < 0.001. ( B ) Examples of telomere abnormalities from ( A ) observed in a telomere FISH assay (upper panel). The incidence of telomere abnormalities in cells lacking ESCO1 or ESCO2 is shown in the bottom panel. ( C ) Sankey diagram showing KEGG pathway analysis of high-confidence proteins associated with ESCO2. Pathway enrichment was performed based on high-confidence ESCO2-interacting proteins, and the results are visualized as a Sankey diagram to illustrate the functional distribution across different KEGG pathways. ( D ) Chord diagram showing KEGG pathway analysis of DNA replication- and repair-related pathways and their associated proteins. Diagram visualizes the relationship between key proteins and their corresponding pathways involved in DNA replication and repair. ( E ) Selected lists of ESCO2-associated proteins analyzed by mass spectrometry. ( F ) Western blot performed to determine ESCO2 and BLM siRNA-knockdown efficiency in HeLa1.2.11 cells. ( G ) Representative images of metaphase telomere FISH in cells from ( G ) (upper panel). The incidence of telomere abnormalities is shown in the bottom panel.

Article Snippet: The antibodies used in this study were: 53BP1 (NB100-904; Novus Biologicals, Littleton, CO, USA), Flag (F3165; Sigma, St. Louis, MO, USA), BLM (A300-110A, Bethyl, Boston, MA, USA), ESCO2 (ab86003; Abcam, Cambridge, UK), Myc (sc-40; Santa Cruz Biotechnology, Santa Cruz, CA, USA), and vinculin (V9131; Sigma-Aldrich, St. Louis, MO, USA).

Techniques: Transfection, Labeling, Staining, Fluorescence, Functional Assay, Mass Spectrometry, Western Blot, Knockdown