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. 2018 Aug 14;46(16):8168–8180. doi: 10.1093/nar/gky721

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© The Author(s) 2018. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 7. — CircRNA are conserved throughout red blood cell aging.(A) Left panel shows a representation of a Percoll-Urografin gradient separation after centrifugation. The immature reticulocyte fraction is found in the top fraction (bellow a neutrophil-rich ring), the lower fractions (F1, F2, F3) separate the aging red-blood cells from young to old (top to bottom). Right panel shows a flow-cytometry measurement of the fractions for CD71 expression and nucleic acid (thiazole orange) staining (representative of 4 donors). (B, C) Graph showing the percentage of CD71+ (B) and thiazole orange + (C) in the fractions from (A) (n = 4). (D) RNA extracted from the fractions in (A) were reverse transcribed and circRNA and linear RNA detection was performed using PCR. 18S and Beta-actin (ActB) were used as a linear control. (D) is representative of 3 donors.