Fig. 3: Nrp1 upregulates the Nfkb1 and Smad3 pathway, suppressing Etv6 and Acox3 expression, leading to reduced levels of OXPHOS and TCA in TECs.

A Transcription factor activity analysis heatmap of DT cells. B Western blotting analysis showing the changes in Nfkb1, Smad3, α-SMA and Pdgfrb after IR and Nrp1 knockout (n = 3 per group). C Dotplot of Tnfa, Nfkb1 and Smad3 in DT cells. D T-SNE plots showing the expression region of Nrp1 and transcription factor activity region of Nfkb1 and Etv6 in DT cells. E The amplification levels of Etv6 after binding with the anti-Nfkb1 monoclonal antibody using qPCR (n = 3 per group). F Dotplot of crotonyl-CoA-producing enzymes in kidney. G A schematic diagram illustrating the enzymes involved in the production of crotonyl-CoA. H Western blotting analysis showing the changes in crotonylation lysine modification after I-R and Nrp1 knockout. I Heatmap displaying the Kyoto Encyclopedia of Genes and Genomes (KEEG) pathways enriched in proteins with changing crotonylation modification sites before and after IR treatment, as well as before and after Nrp1 knockout. J Downregulated KEGG pathways in Nrp1 + DT cells compared to Nrp1-DT cells in scRNA-seq data. The statistical analyzes were two-sided and adjustments were made in P value. K Dotplot of genes related to mitochondrial functional status in DT cells. L Immunofluorescent staining of MitoTracker in primary renal tubular epithelial cells (pTECs) treated with TNF-α and Tmx. M Energy map showing increased oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) in pTECs after Nrp1 knockout (n = 5 per group). Data are representative of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 as determined by one-way ANOVA. Scale bar, 50 μm. Data represent mean ± SEM. Source data are provided as a Source Data file.