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. 2014 Sep 1;28(17):1885–1899. doi: 10.1101/gad.246819.114

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© 2014 Sexton et al.; Published by Cold Spring Harbor Laboratory Press

This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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Figure 3. — TERT knockout phenocopies TPP1 ΔL time course of failures in telomere maintenance and cellular renewal. (A) Targeting schematic for ZFN-mediated integration of a STOP cassette into the endogenous TERT locus. (B) TRAP assay of whole-cell extracts from independent TERT^+/− and TERT^−/− cell lines (n = 3 each) using 333 ng of total protein. (IC) Internal control. (C) Telomere restriction fragment length assay of TERT^+/− and TERT^−/− cell lines over a time course of continuous passaging. (D) Quantification of telomeric repeat hybridization signal lengths in TERT^+/− and TERT^−/− cell lines over a time course of continuous passaging, including those from B. (E) IF of late time point TERT^+/− and TERT^−/− cell lines stained against OCT4. Bar, 200 μm. Dashed square in the top panels indicates an individual nucleus for each genotype that is shown magnified in the bottom panel. (F) Box plot of quantification of nuclear blebbing of late time point TERT^+/− and TERT^−/− cell lines.