Extended Data Fig. 7: AFABP interacts with Transferrin to hinder ISC differentiation and gut regeneration in Drosophila.
From: Obesity impairs gut repair via AFABP-mediated iron overload in intestinal stem cells
a, A diagram outlining the process of using immunoprecipitation coupled with mass spectrometry (IP-MS) to uncover interaction partners of hAFABP in hemolymph. Created by Figdraw (www.figdraw.com). b, DMFOLD, a DeepMSA2 based protein folding model, indicates a strong binding affinity between hAFABP and hTSF1. c-f, Ratio of Smurf (+) files (c, n = 6, 7 independent experiments), deposits (d, n = 20, 23 fields of view for each group), ratio of homeostasis state (e, n = 6, 6 independent experiments), and midgut length (f, n = 21, 20 midguts respectively) from flies fed NCD of indicated genotypes indicated boosting dTSF1 expression in Drosophila fat bodies enhanced intestinal functions in hAFABP-overexpressing flies (control) after gut injury (REC-3D). Overexpression of dTSF1 lines (dTSF1OE) was used. g-i, Ratio of NRE-lacZ+ cells to esg-GFP+ cells (g, n = 11-12 midguts for each group), esg-GFP+ cells to DAPI+ cells (h, n = 12 midguts for each group), and Dl+ cells to DAPI+ cells (i, n = 12 midguts for each group) from flies with indicated treatments and genotypes. RNAi line against dTSF1 (dTSF1RNAi) was used. LUCRNAi as control. j, Relative mRNA expression of dTsf1 genes from indicated genotypes (LUCRNAi, dTSF1RNAi), n = 3 independent experiments. LUCRNAi as control. k, Relative mRNA expression of dTsf1 genes from indicated genotypes (LUCOE and dTSF1OE), n = 3 independent experiments. l, Ratio of esg-GFP+ cells to DAPI+ cells from HFD-induced obese flies treated with vehicle, BPS, DFO, or FAC. Flies treated with NCD only were used as a control (n = 12, 15, 13, 14, 19 midguts respectively). BPS, DFO (two iron chelators), and FAC (an iron supplement) were used. m, Heatmap depicting varied expression patterns of key genes regulating iron homeostasis from NW-WT, OB-AFABPAKO, and OB-WT groups. n, Atomic Absorption Spectroscopy (AAS) analysis of iron content from colonic crypts of indicated treatments and genotypes mice (n = 4 mice). NW-WT as control. Error bars indicate mean +/- SDs. Significance: ns signifies p > 0.05. Unpaired two-tailed Student’s t-tests for (c-f, j, k), one-way ANOVA with Dunnett multiple comparison test for other results. Three independent experiments were performed.
