Fig. 4: Overnutrition alters intestinal stem cell differentiating direction via InR signaling pathway.
a Diagram of the experimental procedure of fly treatment. b–e Immunofluorescence images of midgut with esg-lacZ (red), NRE-GFP (green), and Pros (white) staining from control flies (esg-lacZ/+; NRE-GFP/+) cultured in NF (b), flies cultured in HSD (c), chico+/− flies cultured in NF (d) and chico+/− flies cultured in HSD (e). f, g Quantification of the number of esg+ cells (f) and the ratio of Pre-EE cells to esg+ cells per ROI (g) with indicated genotypes and food treatments of midguts in experiments (b-e). h The percentage of differentiated cells and esg+ cells to total cells in indicated genotypes and food treatments of midguts in experiments (b–e). i, j Quantification of the number of esg+NRE-Pros- ISCs (i) and NRE+ EBs (j) per ROI with indicated genotypes of midguts in young Drosophila. k The ratio of Pre-EE cells to esg+ cells per ROI with indicated genotypes of midguts in young Drosophila. l, m Quantification of the number of esg+NRE-Pros- ISCs (l) and NRE+ EBs (m) per ROI with indicated genotypes of midguts in old Drosophila. n The ratio of Pre-EE cells to esg+ cells per ROI with indicated genotypes of midguts in old Drosophila. o The percentage of differentiated cells and esg+ cells to total cells in indicated genotypes of midguts in old Drosophila. DAPI-stained nuclei (blue). ROI size 84,100 μm2. Yellow asterisks indicate esg+NRE-Pros- ISCs. Yellow arrows indicate esg+Pros+ Pre-EE. Scale bar, 25 μm. For dot plots, bars are mean ± SD. For box plots, box shows median, 25th and 75th percentiles and whiskers represent minima and maxima. Each group was compared to the control group in (i–o). Statistics were measured by one-way ANOVA. Each dot represents one ROI in (f, g) and (i–n). From left to right, n = 22, 19, 21 and 18, respectively in (f–h); n = 18, 19, 35, 23, 16, 17, 17 and 16, respectively in (i–k); n = 20, 31, 15 and 16, respectively in (l–o). Source data are provided as a Source Data file.
