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. 2021 Feb 22;4(4):e202000764. doi: 10.26508/lsa.202000764

Figure S10. R-loops remained unaltered in ageing C9ORF72 spinal MNs.

Figure S10.

Nuclear R-loop foci (in green) and the DNA double-strand break marker 53BP1 (in red) were revealed over a time course from D14 to 80 by IF staining in Hoechst-positive nuclei (in blue) of MAP2-positive (in white) neurons. Refers to Fig 8B. Note the constant R-loop foci count in all Ctrl and C9 lines (green arrowheads) over time, whereas DNA double-strand break foci suddenly increased only in C9 and C9-KO beyond D50 and in WT-KO beyond D60 (red arrowheads). Disappearance of R-loop foci in nuclei upon treatment with RNAse H at D14 (top image row) served as negative control to confirm the specificity of the immunostaining. Arrowheads point only to arbitrary examples. Scale bars = 10 μm.