Extended Data Fig. 5: Multiple sources of Bnl individually contribute to regenerative ISC proliferation in the adult Drosophila midgut, independently of tracheal remodelling.
a, Confocal images of FGF/bnl reporter expression bnl > GFPnlacZ (green) in Sucrose and Pe treated midguts stained with CBP (magenta) to visualize all tracheal tissue and PH3 (magenta). Dotted boxes in left panels indicate the area magnified in the right panels. Scale bars: 100 µm (main figure); 40 µm (close up view). b, bnl > GFPnlacZ staining intensity relative to background within TTCs in midguts as in (a). Two-tailed, unpaired T-test (****P < 0.0001); n = number of TTCs from 12 midguts per condition, indicated in panel. c, d Tracheal coverage (c) and PH3+ ISCs (d) from wild-type (control) animals or upon bnl overexpression within TTCs. Two-tailed, unpaired T-test (****P < 0.0001); n = number of midguts, indicated in panels. e, Bright-field images from Sucrose or Pe treated midguts from wild-type (control) animals or following RNAi-driven bnl knockdown (bnl−IR) within ISCs/EBs. Scale bar: 50 µm. f, g, Tracheal branching (f) and PH3+ ISCs (g) in midguts as in (e). Two-way ANOVA followed by Sidak’s multiple comparisons test (****P < 0.0001); n = number of midguts, indicated in panels. h, Bright-field images from Sucrose or Pe treated midguts from wild-type (control) animals or upon bnl knockdown (bnl−IR) within ECs. Scale bar: 50 µm. i, j, Tracheal branching (i) and PH3+ ISCs (j) in midguts as in (h). Two-way ANOVA followed by Sidak’s multiple comparisons test (****P < 0.0001); n = number of midguts, indicated in panels. k, n, Bright-field images of midguts from wild-type (control) animals or upon bnl overexpression (bnl) within ISCs/EBs (k) or ECs (n). Scale bar: 50 µm. l, m, o, p, Tracheal coverage (l, o) and PH3+ ISCs (m, p) from midguts as in (k, n). Two-tailed, unpaired T-test; n = number of midguts, indicated in panels. l, (***P = 0.0002). m, (**P = 0.0066). o, (*P = 0.0213). p, (**P = 0.0084). Box plots represent maxima, minima and mean. Mean value is indicated on top of boxes. Otherwise, values represent mean ± s.e.m.
