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Fig. 5 Ferroptosis hinders the digestive organ expansion.a, b KEGG pathway analysis of differentially expressed genes in hsd17b12a?/? compared to WT at 3 dpf and 4 dpf (p-value < 0.05). c Single-cell sequencing analysis of the liver, intestine, and pancreas in hsd17b12a?/? and WT at 4 dpf. The ?Liver? label indicates liver-origin cells, the ?Pancreas? label indicates pancreas-origin cells, and the ?Intestine? label indicates intestinal-origin cells. d The proportions of each cell type in hsd17b12a?/? and WT. e KEGG pathway analysis of differentially expressed genes in the intestine of hsd17b12a?/? compared to WT (p-value < 0.05). f Detection of ROS (reactive oxygen species) in WT and hsd17b12a?/? at 4 dpf (n = 6). The white arrowheads indicate the intestine. Scale bar, 0.5 mm. g MDA (malondialdehyde) levels were measured in WT and hsd17b12a?/? at 4 dpf. h RT-qPCR of acsl4a and acsl4b in WT and hsd17b12a?/? at 3 dpf and 4 dpf. i, j Assessment and quantification of intracellular Fe2+ levels in larvae using FerroOrange probe in WT and hsd17b12a?/? at 4 dpf (n > 3). k, l Evaluation of GPX4 protein levels in WT and hsd17b12a?/? by western blot and quantification of GPX4 relative to GAPDH (n = 3). m Immunofluorescence analysis of GPX4 protein in the primitive intestine of WT and hsd17b12a?/? at 4 dpf (n = 3). The white arrows indicate the intestinal epithelia. Scale bar: 25 ?m. n Fluorescence imaging of the liver and exocrine pancreas in hsd17b12a?/? treated with the Acsl4 inhibitor Rosi (rosiglitazone) (n > 3). o Measurement of the exocrine pancreas and liver areas in both untreated and Rosi-treated hsd17b12a?/?. Scale bar: 250 ?m. p, q Fluorescence imaging and quantification of the exocrine pancreas and liver areas in WT, hsd17b12a?/?, and Fer-1 (Ferrostatin-1)-treated hsd17b12a?/? at 5 dpf (n > 3). Scale bar: 200 ?m. r RT-PCR of fabp10a and trypsin at 5 dpf after treatment with Fer-1. Data in g, h, j, l, o, q, r are represented as mean ± S.D., using a two-tailed Student?s t test; each point represents an independent biological sample (n ? 3). Source data are provided as a Source Data file.