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. 2018 Aug 29;13(8):e0201787. doi: 10.1371/journal.pone.0201787

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© 2018 Kordyukova et al

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Fig 1 — (A) RT-qPCR analysis of RNA immunoprecipitated with anti-HA from ovary lysates of nosGal4; UAS-HeT-A-HA; UAS-spnE_sh flies and control nosGal4; UAS-spnE_sh flies. Fold changes (FC) for RNA enrichments in RNA IP from ovaries of HeT-A-HA transgenic strain versus control are shown. rp49 was used for normalization. The error bars represent standard deviations (SD) of 2 biological replicas. Western blot analysis of immunoprecipitated HeT-A Gag-HA is shown to the right. The antibodies used for Western blotting are indicated to the right. (B-G) RNA FISH combined with immunostaining was performed on ovaries of nosGal4; UAS-HeT-A-HA; UAS-spnE_sh flies. (B) HeT-A RNA (green) colocalizes with HeT-A Gag-HA (red) in germarium. Enlarged region (white rectangle) is shown. HeT-A spheres (arrows) are observed in cystoblasts. (C) HeT-A RNA (green) colocalizes with HeT-A Gag-HA (red) in nurse cell nuclei. A fragment of a stage 7 egg chamber is shown to the left. An enlarged nucleus of a germline nurse cell (white rectangle) is shown. (D) HeT-A RNA FISH (green) combined with HeT-A Gag-HA (red) and HOAP (magenta) immunostaining in a nurse cell nucleus from a stage 7 egg chamber is shown. Telomeric localization of HeT-A RNP is shown by arrows. (E) HeT-A RNA (green) and HeT-A Gag-HA (red) form large aggregates (arrows) in the cytoplasm of nurse cells. A fragment of a stage 5 egg chamber is shown. For an entire egg chamber see Part A in S2 Fig. (F) TART transcripts (green) are co-localized with HeT-A Gag-HA (red) in the nurse cell cytoplasm (arrows). A fragment of a stage 7 egg chamber is shown. (G) TART RNA (green) colocalization with telomere-specific protein HOAP (magenta) is shown (arrows). HeT-A Gag-HA (red) partially colocalizes with TART RNA in nurse cell nuclei. Image of an individual nurse cell nucleus from a stage 6 egg chamber.