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Line G04609 enhancer trapping evaluation: genomic mapping and expression of neighboring genes (midline and H15).

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posted on 31.07.2014, 03:19 authored by Stefan Koelzer, Yvonne Kölsch, Kristen A. Panfilio

(A) Genome browser views (version Tcas_3.0) show the insertion site (red) within the context of the entire chromosomal linkage group (ChLG, grey background) and enlarged for the region 75 kb 3′ to the insertion site (white background). Predicted genes (Tcas_3.0 official gene set) within this region are labeled and shown in green (dark green: entire gene, light green: exons). Blue bars span the exons used as cDNA template for in situ hybridization. In the case of H15, this spanned the TC013511 and TC013513 gene models, omitting the linking region marked by the thin line. In newer models, Tc-mid is represented by au2.g7325.t1 or au3.g6464.y1; Tc-H15 is au2.g7328.t1 or au3.g6465.t1 (explained in Table 2). (BJ) In situ hybridization for the nearby candidate genes Tc-mid (B–F) and Tc-H15 (G–J), for the early stage of serosal window closure (B,C,G,H), and for the later stages of the retracted germband and serosal contraction/dorsal organ stage (D–F,I–J). See the main text for descriptions of these expression domains. All images labeled with the same letter are of the same embryo (except D3). Letter-prime images show the nuclear counterstain. Views are lateral (B,C,D1,F,I1,J), ventral (D2,D3,G,H,I2,I3), or dorsal (E), with anterior left. In D, the serosa has been removed from the embryo, exposing the yolk (Y). Annotations: red dashed line, serosal window outline (G,H); arrowheads, the outer, folded parts of the contracting serosa (F,I1,J). Additionally, dashed outlines in inset images (D3,I3) outline the anterior margin of the legs. Sense strand controls (C,F,H,J) show increased background levels in the folded, thickened outer regions of the contracting serosa (F,J), and non-specific stain in the pleuropodia (arrows in F,J). Abbreviations as in previous figures, and Em, embryo. Scale bars are 100 µm, except for 50 µm for the inset images (D3,I3).