Fig. 2: Aging alters intestinal stem cell differentiating direction in Drosophila.

a A model of cell lineage in Drosophila midgut epithelium and the characteristic marker for each cell type was indicated. b–d Representative immunofluorescence of midguts with GFP (green), lacZ (red), and Pros (white) staining from 10d (b), 30d (c), and 50d (d) flies (esg-lacZ/+; NRE-GFP/+). e, f Quantification of the number of esg⁺ cells (e) and esg⁺Pros⁺ cells (f) per region of interest (ROI) of midguts in experiments (b–d). g The percentage of ISCs, EBs, and Pre-EEs to esg⁺ cells of midguts in experiments (b–d). h The percentage of differentiated cells and esg⁺ cells to total cells of midguts in experiments (b–d). i The percentage of ECs and EEs to total differentiated cells of midguts in experiments (b–d). j, k The relative mRNA expression level of EC-specific genes (j) and EE-specific genes (k) from whole midgut in old flies compared with young flies. The mRNA expression level of each gene from young flies was normalized to 1. Each experiment was repeated for 3 times. ROI size 84,100 μm2. DAPI-stained nuclei (blue). Yellow asterisks indicate esg⁺NRE^-Pros⁻ ISCs. Yellow arrows indicate esg⁺Pros⁺ Pre-EE. Scale bar, 25 μm. For dot plots and (g), bars are mean ± SD. For box plots, box shows median, 25th and 75th percentiles and whiskers represent minima and maxima. Statistics were measured by one-way ANOVA in (e–i) and two-tailed, unpaired Student’s t-test in (j, k). Each dot represents one ROI in (e, f). n = 23, 30 and 15 in 10d, 30d, and 50d group, respectively in (e–i). Source data are provided as a Source Data file.