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

Figure 3. Notch expressing cells in first instar lymph gland of Drosophila are multipotent.

(A) N-GFP expression in a first instar lymph gland. N expression is detectable only till late first instar (also see Figure 1[E, M–O]). (B–E") shows that activation of G-TRACE with N Gal4, in a short window (following scheme in B) results in labeling of majority of the cells of third instar lymph gland. N lineage traced cells (green; [C–E"]; n = 15) include both crystal cells ([D'-D"], hnt, grey; n = 10) and plasmatocytes ([E'–E"], P1, grey; n = 10). (F) N expression is reduced to 2 cells by about 18 hr AEH (n = 7). (G–H) show that restrictive lineage tracing of one (H; n = 6) or two (G; n = 8) HSCs result in a clonal propagation, which spans through the entire horizontal length of the gland. (I–J) Each of these cells is equipotent in giving rise to both plasmatocytes ([I], P1, grey, n = 9) and crystal cells ([J], hnt, grey; n = 9). Scale bar = 5 μm for Figure 3A and F, for rest is 20 μm Genotypes are shown on top of corresponding panels. DAPI marks the nucleus. Hours after larval hatching are as indicated in each panel. Also see Figure 3—figure supplement 1–2.

Figure 3.

Figure 3—figure supplement 1. Fate map of the progenitors arising from Notch expressing HSCs.

Figure 3—figure supplement 1.

(A) shows dome-GFP expression in progenitors of a late third instar lymph gland (n = 10). (B–C") show that dome expressing progenitors upon lineage tracing (C–C', green) from 18 hr AEH using the scheme in (B) give rise to all the cells of the lymph gland. Red indicates current expression of dome whereas green shows lineage of the same (C–C"; n = 10). (D–E"') show that dome lineage comprises of both crystal cells (D'–D"', Hnt, grey; n = 13) and plasmatocytes (E–E"', P1, grey; n = 15). Genotypes are shown on top of corresponding panels. Scale bar = 20 μm. DAPI marks the nucleus. Hours after larval hatching are as indicated in each panel.
Figure 3—figure supplement 2. Notch requirement in crystal cell maintenance at a later stage in a third instar lymph gland.

Figure 3—figure supplement 2.

(A–B") shows that, upon lineage tracing beyond HSC lifeline (following scheme in A), Notch lineage (green, [B, B"]) comprises of only crystal cells (red, live expression, [B, B'], n = 10). (C) Notch expression (green) in a wild-type late third instar lg overlaps with crystal cells (Red, Hnt, n = 10). Scale bar is 20 μm. Genotypes are shown on top of corresponding panels. DAPI marks the nucleus. Hours after larval hatching are as indicated in each panel.