Fig. 3: Differential localisation detection levels of mOr2-Piwi1 and GFP in a piwi1mOr2/P2A-GFP CRISPR-KI double reporter line.
From: A population of Vasa2 and Piwi1 expressing cells generates germ cells and neurons in a sea anemone
A–D’ Confocal images of double transgenic piwi1mOr2/P2A-GFP polyps immunostained for mOr2 (A–D; yellow) and GFP (A’–D’; cyan). Cross sections of a female gonadal mesentery show high levels of mOr2 and GFP that colocalize to basiepithelial cells in the reticular tract (A–C’; arrowheads), and to oogonia (pink arrowheads) and developing oocytes (‘o’) in gonadal regions (A-A’, C-C’). Note cytoplasmic restriction of mOr2-Piwi1 in basiepithelial cells (B) and to perinuclear granules in oocytes (C; nuage). Extension of GFP localization to the cell nucleus (B-C’; ‘n’) validates the successful GFP-Piwi1 separation and the use of the piwi1P2A-GFP allele for lineage tracing. Low levels of GFP and mOr2 colocalize to cells at the basis of the cnidoglandular tract (A-A’; arrows at ‘cgt’), retractor muscle (A-A’; arrow at ‘rm’), and parietal muscle (A-A’, D-D’; arrows at ‘pm’) of the adult mesentery. Grey: Hoechst DNA dye. cgt cnidoglandular tract, ct ciliated tract, ep epidermis, it intermediate tract, m mesentery, n nucleus, o oocyte, pm parietal muscle, rm retractor muscle, rt reticular tract, sg somatic gonad. Scale bars: 50 µm (A) and 10 µm (B–D). All experiments were performed twice with similar results.
