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. 2016 Oct 26;5:e18295. doi: 10.7554/eLife.18295

Figure 5. Hierarchical relationship between dome and Notch expressing cells in the first instar lymph gland.

(A) Show the scheme of N lineage tracing followed throughout this panel. (B) Schematic representation of predicted cell types arising from N lineage tracing in A. A red cell indicates live expression of N. A green cell indicates that N was active in this or its precursor cell but is not active now. A yellow cell indicates continuous activation of N>GTRACE indicating that this cell expressed N earlier and is still expressing. A magenta cell indicates dome-MESO expression. (C–C'") First instar lymph gland showing mutually exclusive yellow (both N live and lineage traced expression; stars) and magenta (dome-MESO-EBF2) cells at 8 hr (n = 6). (D–D''") also shows few N expressing cells (yellow; stars in D–D''") that do not express dome-MESO-EBF2 (magenta; D'"–D''"; n = 7) at 15 hr. (E–E''") At 18 hr, some N expressing cells (arrows; E–E''"; n = 6) also express dome (arrows; E"–E''"), while some do not (stars; E–E''"), indicating a possible differentiation at this time point. (F–F'") Second instar early (30 hr AEH) lymph gland does not actively express N (absence of red live expression in F) but contains N lineage traced cells (green; F', F"'; n = 5), which are now also expressing dome-MESO (magenta; F"–F'"). Also noteworthy are few cells (arrowheads) that are not lineage traced for N (not green) but still express dome (arrowheads, D–F'"). (G–G'") dome-MESO-EBF2 (magenta; G, G"–G'") expression in restricted N lineage (green; G'–G'") traced lymph gland following scheme in A. Scale bar is 20 μm for G–G"' for rest it is 5 μm. Genotypes are shown on top of corresponding panels. TOPRO marks the nucleus.

Figure 5.

Figure 5—figure supplement 1. Lineage tracing in an identical window reveals the hierarchical relationship between dome and Notch expressing cells.

Figure 5—figure supplement 1.

(A–B) Expression of Ci (Cubitus interruptus, red: B–B"), a marker of progenitors in transient N lineage traced (green) lymph gland following the scheme in A (n = 10). (C) is a cartoon showing overlap of dome and Ci (yellow), both validated markers of progenitors (D–D") Following restricted lineage tracing scheme in A, dome lineage tracing (green) marks much less number of progenitors (Ci; red; n = 5) in comparison of similarly lineage traced N. Compare D–D" to B–B". (E–E') Overlap of dome-MESO and tepIV, a downstream responder of same JAK/STAT pathway in a third instar lymph gland. (F–H) show three examples of restricted labeling of tepIV, following lineage tracing scheme in A (green; n = 8), marking much less number of cells in comparison of similarly lineage traced N (compare F–H with B). Scale bar 20 μm. Genotypes are shown on top of corresponding panels. TOPRO marks nucleus in E–E', while DAPI marks nucleus for rest.