Pilocytic Astrocytomas Have Telomere-Associated Promyelocytic Leukemia Bodies without Alternatively Lengthened Telomeres Tania Slatter, Jennifer Gifford-Garner,

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Pilocytic Astrocytomas Have Telomere-Associated Promyelocytic Leukemia Bodies without Alternatively Lengthened Telomeres Tania Slatter, Jennifer Gifford-Garner, Anna Wiles, Xin Tan, Yu-Jen Chen, Martin MacFarlane, Michael Sullivan, Janice Royds, Noelyn Hung The American Journal of Pathology Volume 177, Issue 6, Pages 2694-2700 (December 2010) DOI: 10.2353/ajpath.2010.100468 Copyright © 2010 American Society for Investigative Pathology Terms and Conditions

Figure 1 Grade 1 astrocytomas are positive for APBs. A: Measurement of APBs in astrocytomas. APBs (indicated with an arrow) had co-localized PML (green) and telomere DNA (red) fluorescent signals in nuclei (not shown) on analysis of paraffin-embedded samples at ×630 magnification. The criteria for an APB-positive cell included at least two co-localized areas greater than 0.9 μm in size (profile, bottom). Scale bar = 20 μm. B: Distribution of APB measurements among grade (G) 1−4 astrocytomas. The percentage of APB-positive cells is plotted against tumor grade. The criteria for APB-positive tumors included APBs greater than 0.5% of cells, as indicated by a dashed line. The median APB measurement is represented with a solid horizontal bar. The American Journal of Pathology 2010 177, 2694-2700DOI: (10.2353/ajpath.2010.100468) Copyright © 2010 American Society for Investigative Pathology Terms and Conditions

Figure 2 Grade 1 astrocytomas are ALT negative by telomere length analysis. A: Measurement of ALT in astrocytomas by TRF length. Determination of mean TRF lengths followed standard procedures for the TRF assay using agarose gel electrophoresis. A TRF profile with long telomeres (mean TRF length >16 kb) that typically ranged widely in length (approximately <3–> 50 kb) was classified as ALT positive, and was present in grade (G) 2−4 tumors (highlighted by asterisks). Shorter telomeres with less length variation were classified as ALT negative, and presented in grade (G) 1 and 4 astrocytomas. Molecular weight marker, kb. B: Distribution of mean TRF length measurements among grade (G) 1−4 astrocytomas. The mean TRF length is plotted against tumor grade. The criteria for an ALT-positive status included a mean TRF length above 16 kb, as indicated by a dashed line. The median TRF measurement is represented with a solid horizontal bar. The American Journal of Pathology 2010 177, 2694-2700DOI: (10.2353/ajpath.2010.100468) Copyright © 2010 American Society for Investigative Pathology Terms and Conditions