Journal: Nature Communications
Article Title: Renal insulin-like growth factor binding-protein 7 is a critical promoter of progressive diabetic kidney disease
doi: 10.1038/s41467-025-66490-5
Figure Lengend Snippet: A KEGG enrichment analysis of upregulated (above) and downregulated (down) genes following IGFBP7 overexpression (P adj. Value, false discovery rate). B GSEA plots of fatty acid metabolism gene sets. C KEGG module analysis of fatty acid metabolism. The results showed a significant enrichment in the fatty acid β-oxidation pathway (P adj. Value, false discovery rate). D GSEA plots for oxidative phosphorylation and TCA cycle genes. E Real-time PCR analysis of the mRNA expression of electron transport chain (ETC, Complex I–V) complex genes in BP7-Tg and WT kidney at 8 months of age ( n = 6, two-tailed unpaired Student’s t test). F Western blot of PGC-1α in FF versus cKO mice ( n = 6, one-way ANOVA with Tukey’s multiple comparisons test). G Real-time monitoring the oxygen consumption rate (OCR) in HK2 with or without IGFBP7 overexpression (BP7-EV ( n = 3), BP7-OE ( n = 4); biological replicates). H Real-time monitoring the OCR in HG-treated HK2 with or without IGFBP7 knockdown ( n = 3). I Relative OCR of HG-treated HK2 with or without IGFBP7 overexpression ( n = 12, two-tailed unpaired Student’s t test). J Etomoxir-induced decreases in OCR from HG-treated HK2 ( n = 12, two-tailed unpaired Student’s t test). K Schematic of the mechanism by which IGFBP7 induces impaired fatty acid degradation and lipid accumulation in renal tubular epithelial cells; key components and inhibitors involved are shown (by FigDraw 2.0). L Schematic of in vitro co-culture experiments. M Western blot analysis of Wilms’ Tumer 1 (WT-1) and Nephrin protein levels in MPC5 cells under mTEC conditioned media ( n = 4, one-way ANOVA with Tukey’s multiple comparisons test). N Western blot analysis of COL-1 and αSMA protein levels in NIH-3T3 cells under mTEC conditioned media ( n = 4, one-way ANOVA with Tukey’s multiple comparisons test). O Kidney spheroids from FF and IGFBP7-cKO mice. P Western blotting and quantification of COL-1 and αSMA in FF and cKO mouse kidney spheroids treated with high glucose ( n = 3, one-way ANOVA with Tukey’s multiple comparisons test). Data represent the mean ± SEM. n.s. not significant, FF floxflox, cKO conditional knockout in tubular, WT wild type, BP7-Tg IGFBP7 overexpression transgenic, HG high glucose, EV empty vector, OE overexpression. Source data are provided as a Source Data file.
Article Snippet: Antibodies specific for IGFBP7 (ab171085, ab214262: Abcam; and 19961-1-AP: Proteintech), COL-1 (66761-1-Ig; Proteintech), αSMA (14395-1-AP; Proteintech), STAT3 (10253-2-AP; Proteintech), pSTAT3 (R381552; Zen-Bio), AcSTAT3 (#2523; CST), STAT1 ( R25799 ; Zen-Bio), STAT5 (R381427; Zen-Bio), PGC-1α (R381615; Zen-Bio), pAMPK (AP1441; Abclonal), AMPK (A1229; Abclonal), H3K4Me1 (A22079; Abclonal), H3K4Me3 (A22146; Abclonal), H3K27Ac (A22264; Abclonal), Histone3 (A2348; Abclonal), Plin2 (15294-1-AP; Proteintech), KIM-1 (30948-1-AP; Proteintech), NLRP3 (WL02635; Wanlei), TNFα (60291-1-Ig; Proteintech), γH2AX (R381558; Zen-Bio), WT-1 (ab267377; Abcam), Nephrin (ab216341; Abcam), NFκB1 (10745-1-AP; Proteintech), HSPA5 ( R24618 ; Zen-Bio), SIRT2 (ET1611-72; HuaBio), Flag (20543-1-AP; Proteintech), HA (51064-2-AP; Proteintech), Myc (60003-2-Ig; Proteintech), GFP (M20004; Abmart), MDM2 (66511-1-Ig; Proteintech) and β-actin (bs-10966R; Bioss).
Techniques: Over Expression, Phospho-proteomics, Real-time Polymerase Chain Reaction, Expressing, Two Tailed Test, Western Blot, Knockdown, In Vitro, Co-Culture Assay, Knock-Out, Transgenic Assay, Plasmid Preparation