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. 2018 May 7;215(5):1463–1480. doi: 10.1084/jem.20170997

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© 2018 Barbieri et al.

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Figure 2. — Irradiation promotes active retrotransposition in HSCs. (A) The huL1-GFP transgene and position of the primers used to detect retrotransposition. (B–D) Experimental design and analysis of L1 retrotransposition by Taqman (B and D) or FACS (C) in HSCs and progenitors of L1-GFP mice before (NIR; B–D) or after TBI (IR; B and C). (B) Means ± SEM, n = 3 (WT), 7 (L1-GFP NIR), and 13 (L1-GFP IR) mice from three independent experiments. Mann-Whitney test. (C) Each triangle represents an individual mouse. Means ± SEM from two independent experiments. Mann-Whitney test. (D) n = 7 (HSC, CMP, and GMP) and 6 (MEP) mice from two independent experiments; ANOVA with Dunnett’s multiple comparison test. (E) Kinetics of L1 retrotransposition after TBI by Taqman-based qPCR assay. Left, experimental design; middle, representative FACS images of HSC sorting just after TBI; right, GFP expression normalized on 5S rDNA. Means from two independent cultures. Open circles, NIR; closed squares; IR. (F) qPCR analysis of the number of genomic copies of L1 ORF2 in HSCs 1 mo after TBI (IR) or not (NIR), normalized to 5S rDNA. n = 6 (NIR) and 8 (IR) mice. Means ± SEM from two independent experiments. Mann-Whitney test. *, P < 0.05; ***, P < 0.001.