Fig. 3: Heterogeneity of progenitors and dorsal branch. | Nature Communications

Fig. 3: Heterogeneity of progenitors and dorsal branch.

From: A single-cell atlas of Drosophila trachea reveals glycosylation-mediated Notch signaling in cell fate specification

Fig. 3

a Dotplot showing the expression of exp and kni in each cluster. b Schematic diagram showing the tracheal system of a pupa. The red boxed region indicates DB. c–e Dependence of DB on Exp and Kni. Images of DB in control (left), expRNAi (middle) and kniRNAi (right) flies. Each experiment was repeated independently for three times with similar results. f Dotplot showing the expression of bs, mirr, bru2, and wun in each cluster, with a significant enrichment in PC. g Scatter plot showing the number of incorporated EdU foci in tracheal progenitors of control (n = 18), bsRNAi (n = 11), mirrRNAi (n = 12), wunRNAi (n = 10), bru2RNAi (n = 8) and ctRNAi (n = 6) pupae. h Bar graph plotting the velocity of migrating progenitors (n = 3). g, h Three biologically independent replicates were performed for each experiment. Data are presented as mean values ± SD. An unpaired two-tailed t-test was used for all statistical analyses. No adjustments were made for multiple comparisons. i UMAP plot representing subclusters of PC. j Heatmap showing the expression of the unique genes for each cluster. k, l Cut expression in progenitor cells. k cut-expressing progenitors were visualized by ct-Gal4-driven UAS-GFP expression. PCs are indicated by a dashed line. l Cut expression was through immunostaining of Cut in the dissected trachea of P[B123]-RFP-moe flies. m–m”’, The expression of Cut and InR-SPARK in PCs of L3 larvae. m Nuclei of progenitors are indicated by DAPI staining. m’ The presence of cut-expressing progenitors and progenitors with high insulin receptor activity as indicated by GFP droplets. Arrowhead points to progenitors expressing Cut while exhibiting low InR activity. m” PCs are marked by P[B123]-RFP-moe. m”’ Merge image. k–m”’ Each experiment was repeated independently with similar results for three times. DT dorsal trunk, DB dorsal branch, TC transverse connective, ASP air sac primordium, VB visceral branch, SB spiracular branch, PC progenitor cells, LT lateral trunk, GB ganglionic branches. Scar bars: 200 μm (c–e), 50 μm (k, l), 20 μm (m-m”’). Source data are provided as a Source Data file.

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