Fig. 2: The activities of insulin pathway and Hippo pathway in larval–pupal transition and starvation conditions.

a Volcano plot of RNA-seq showing the comparison of gene expression profiles of tracheal progenitors in L3 larvae and white pupae during larval–pupal transition. b GO analysis reveals top functional clusters among the differentially expressed genes. c Heatmap showing the differential expression of genes in pupae relative to L3 larvae. d Volcano plot of RNA-seq showing differentially regulated genes in tracheal progenitors by starvation. e GO analysis reveals top functional clusters among the differentially expressed genes. f Heatmap showing the differential expression of genes by starvation. g Venn plot of differentially expressed genes (DEGs) in tracheal progenitors from starved L3 larvae and 0 h APF white pupae, compared to control L3 larvae. h–m” Dynamic activation of reporters in L3 larvae (h, i, j, k, l, m), 0 h APF pupae (h’, i’, j’, k’, l’, m’) and L3 larvae post-starvation (h”, i”, j”, k”, l”, m”). h Four independent experiments were repeated with similar results. l–m”, Staining tracheal progenitors of btl > GFP in L3 larvae (l, m), 0 h APF pupae (l’, m’) and L3 larvae post-starvation (l”, m”) with α-lacZ antibodies (magenta). m–m” Merge images. n, o, Merge images showing lnR-SPARK droplets (arrowheads, green) and ex-lacZ (magenta) in L3 larvae (n) and white pupae (o). Scale bars: 50 μm (h–o). p Box plot represents the relative level of ex-lacZ normalized to GFP signal from btl > GFP. 0 h APF (n = 10; p = 1.38e-4) and starvation (n = 11; p = 9.07e-6). n = 15 control flies. Four biologically independent experiments were performed. q Box plot showing the relative signals of SPARK reporters. InR-SPARK (p = 2.02e-9), AMPK-SPARK (p = 9.38e-4) and Akt-SPARK (p = 1.61e-3). n = 16, 12, 10, 6, 5, 4 from the left to the right. Three biologically independent experiments were performed. p, q Unpaired two-tailed t-test was used for all statistical analyses. Results are presented as median with minima and maxima. 25th–75th percentile (box) and 5th ~ 95th (whiskers) are indicated in the box plots. No adjustments were made for multiple comparisons in this figure. Source data for (p, q) are provided as a Source Data file.