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. 2017 Jan 17;7:40372. doi: 10.1038/srep40372

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Copyright © 2017, The Author(s)

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To detect the effect of ERR knockdown on germ/cyst cell lineages in Drosophila testes, we analyzed the localization of bag of marbles (bam; (A,B) in green), vasa (C,D) in green) eyes absent (eya; (E,F) in green) and alpha-spectrin ((G,H), in green) in the control and knockdown testes through immunofluorescence. DAPI (blue) was used to stain the nuclei of the cells. We detected bam in marking the germ cell/spermatogonia in control testes (Panel A; green in CONTROL) but very randomly localised/feeble signals were observed in ERR knockdown testes (Panel B; green in ERR-TRiP). Similarly vasa marking the germ cells were prominently evident in control testes (Panel C; green in CONTROL) showing compactly arranged germ cells underneath the hub, and loosely, yet uniformly scattered in tissue apex. No such discrete pattern could be seen in knockdown testes (Panel D; green in ERR-TRiP) where germ cells marked, were randomly arranged. Eya marking the late somatic cyst cells were optimally evident in control testes (green, Panel E) but very reduced number as well as intensity of eya signals were visible in ERR knockdown testes (Panel F; green in ERR-TRiP) despite the presence of cells as evident in the nuclear staining of the same, through DAPI (blue signals). Similarly, in control testes, alpha-spectrin marked profusely branched fusomes (green; Panel G), indicative of the normal 8–16 celled secondary spermatocytes. In contrast ERR knockdown testes had a very few branched fusomes/more linear and small fusomes (Panel H; ERR-TRiP) as marked by alpha-spectrin.