Journal: Kidney360

Article Title: Arginase-1 Expressing Macrophages Interact with Myofibroblasts to Regulate Insulin-Like Growth Factor-1-Dependent Tubule Repair

doi: 10.34067/KID.0000001150

Figure Lengend Snippet: Igf1 is upregulated in an Arg1 -dependent manner in the outer medulla after kidney injury. Arg1 WT and Arg1mko were subjected to IRI/CL-NX and kidneys harvested and microdissected for analysis on day 2. (A) Igf1 mRNA levels in the outer medulla of healthy kidneys and Arg1mko kidneys, n =8 mice per group (B). Immunofluorescence staining of outer medullary sections of Arg1 WT healthy, Arg1 WT injured, and Arg1mko injured kidneys with phospho-IGF1R (red), LTL (green), and DAPI (blue). (C) MFI of phospho-IGF1R signals as shown in (B) were quantified using ImageJ, with five high power fields/kidney section analyzed/group ( n =4 mice). (D) Analysis of mRNA expression of Igf1 in MACS sorted F4/80 + macrophages; ( n =5 mice). (E) Analysis of mRNA expression of Igf1 by naïve, Arg1 low BMDM (transwell coculture of BMDM with BMDM) versus alternatively activated, Arg1 high BMDM (transwell coculture of BMDM with PCRC+GM-CSF, [ n =3]). (F) IGF1 protein concentrations determined by ELISA in the 24 hours conditioned media collected from the conditions described in (E) showing the secreted IGF1 protein levels in MՓ CM1 (naïve, Arg1 low macrophages) and MՓ CM2 (alternatively activated, Arg1 high macrophages) ( n =3/group). P values are shown, and data are presented as mean±SEM. Arg1, arginase-1; Arg1 mko , macrophage-specific Arg1 null; BMDM, bone marrow-derived macrophages; DAPI, 4′,6-diamidino-2-phenylindole; GM-CSF, granulocyte-monocyte colony stimulating factor; IGF1R, IGF1 receptor; IRI, ischemia-reperfusion kidney injury; IRI-CL/NX, ischemia-reperfusion with contralateral nephrectomy; LTL, Lotus tetragonolobus lectin; MACS, magnetic activated cell sorting; MFI, mean fluorescence intensity; MՓ CM, macrophage-derived conditioned medium; PCRC, primary cultured renal cells; pIGF1R, phosphorylated IGF1R; WT, wild type

Article Snippet: IGF1 ELISA assays were performed using Mouse IGF1 ELISA kit (Cat. # ELM-IGF1-1, RayBiotech), and GH ELISA assays were performed by Human GH ELISA Kit (Cat. # ELH-GH-1, RayBiotech).

Techniques: Immunofluorescence, Staining, Expressing, Enzyme-linked Immunosorbent Assay, Derivative Assay, FACS, Fluorescence, Cell Culture

Journal: Kidney360

Article Title: Arginase-1 Expressing Macrophages Interact with Myofibroblasts to Regulate Insulin-Like Growth Factor-1-Dependent Tubule Repair

doi: 10.34067/KID.0000001150

Figure Lengend Snippet: Igf1 is expressed by outer medullary myofibroblasts and endothelial cells. (A) Igf1 expression is greatest in human kidney myofibroblasts based on scRNA-seq of human kidney biopsy samples from publicly available KPMP datasets. (B) scRNA-seq from healthy mouse kidneys confirms that Igf1 expression is highest in myofibroblasts. (C) Igf1 mRNA levels in MACS sorted PDGFR β - and PDGFR β + cell populations from the outer stripe of healthy and injured kidneys, n =5 mice per group. (D) UMAP visualization of identified cell clusters present in freshly isolated PDGFR β + cells from mouse outer stripe 2 days post-IRI. (E) Heatmap showing Igf1 expression across different clusters with highest expression in endothelial cells and myofibroblasts. (F and G) Igf1 levels in FACS-sorted PDGFR β lo and PDGFR β hi cells (F) and PECAM lo and PECAM hi cells (G) isolated from day 2 IRI kidneys, n =4 mice per group. P values are shown, and data are presented as mean±SEM. IRI, ischemia-reperfusion injury; KPMP, Kidney Precision Medicine Project; PDGFR, platelet derived growth factor receptor; PECAM, platelet endothelial cell adhesion molecule-1; scRNA-seq, single-cell RNA sequencing; UMAP, Uniform Manifold Approximation and Projection

Article Snippet: IGF1 ELISA assays were performed using Mouse IGF1 ELISA kit (Cat. # ELM-IGF1-1, RayBiotech), and GH ELISA assays were performed by Human GH ELISA Kit (Cat. # ELH-GH-1, RayBiotech).

Techniques: Expressing, Isolation, Clinical Proteomics, Derivative Assay, Single Cell, RNA Sequencing

Journal: Kidney360

Article Title: Arginase-1 Expressing Macrophages Interact with Myofibroblasts to Regulate Insulin-Like Growth Factor-1-Dependent Tubule Repair

doi: 10.34067/KID.0000001150

Figure Lengend Snippet: Arg1+ macrophage conditioned media induces myofibroblasts and endothelial cells to produce Igf1 in vitro . (A) scRNA-seq analysis of cultured PCRC cells demonstrates that myofibroblasts increase Igf1 mRNA expression in coculture with WT macrophages. (B) PTEC express the highest level of Igf1r after coculture with macrophages. (C) quantitative PCR analysis shows that cultured MACS-enriched PDGFR β + cells express high levels of Igf1 mRNA in response to culture in MՓ CM2 compared with MՓ CM1 and MՓ CM3, n =5. (D) PDGFR β -enriched cells secrete high levels of IGF1 protein in response to culture in MՓ CM2, n =7. (E and F) HUVEC endothelial cells significantly increase Igf1 mRNA expression (E) and IGF1 protein secretion (F) when treated with MՓ CM2 compared with MՓ CM1 and MՓ CM3, n =5. P values are shown, and data are presented as mean±SEM. HUVEC, human umbilical vein endothelial cells; MAC, macrophages; PTEC, proximal tubular epithelial cells; TEC, other tubular epithelial cells.

Article Snippet: IGF1 ELISA assays were performed using Mouse IGF1 ELISA kit (Cat. # ELM-IGF1-1, RayBiotech), and GH ELISA assays were performed by Human GH ELISA Kit (Cat. # ELH-GH-1, RayBiotech).

Techniques: In Vitro, Cell Culture, Expressing, Real-time Polymerase Chain Reaction

Journal: Kidney360

Article Title: Arginase-1 Expressing Macrophages Interact with Myofibroblasts to Regulate Insulin-Like Growth Factor-1-Dependent Tubule Repair

doi: 10.34067/KID.0000001150

Figure Lengend Snippet: Myofibroblast conditioned media activates the IGF1 receptor and tubular proliferation. (A) Immunofluorescence staining of cultured PCRCs exposed to myofibroblast CM1 (MF CM1, induced by Arg1 low macrophages) or MF CM2 (induced by Arg1 high macrophages) for 15 minutes. 10 ng/ml IGF1 stimulation was used as positive control. The activated IGF1 receptor (green) was detected using anti–phospho-IGF1R antibody and proximal tubule cells identified using AQP1 (red) and nuclei by DAPI staining (blue). (B) Representative densitometry of the images as in (A) showing mean fluorescence intensity, n =4. (C) Representative immunoblot performed on lysates from PCRCs treated with the indicated CM showing pIGF1R, with total IGF1R and GAPDH as loading controls, and (D) corresponding bar graph showing the quantification normalized to total IGF1R, n =3. (E) Ki67 staining of PCRCs exposed to the indicated myofibroblast CM for 24 hours (MF CM3=induced by Arg1 MKO macrophages). 10 ng/ml IGF1 was used as the positive control. (F) Quantification of the percentage of Ki67 positive cells as in (E), n =3 (G) Immunofluorescence costaining of Ki67 and LTL in the outer medulla of WT control kidneys ( Arg1 WT healthy), WT kidneys on day 2 post-IRI ( Arg1 WT Injured), and Arg1 mko kidneys on day 2 post-IRI (Arg1 mko Injured). DAPI staining of nuclei is shown (blue). Arrows indicate Ki67+ nuclei of tubular cells from LTL+proximal tubules. (H) Quantification of numbers of Ki67+ nuclei adjacent to LTL+proximal tubules as in (G), n =5. P values are shown, and data are presented as mean±SEM. GAPDH, glyceraldehyde-3-phopshate dehydrogenase; MF CM, myofibroblast conditioned media.

Article Snippet: IGF1 ELISA assays were performed using Mouse IGF1 ELISA kit (Cat. # ELM-IGF1-1, RayBiotech), and GH ELISA assays were performed by Human GH ELISA Kit (Cat. # ELH-GH-1, RayBiotech).

Techniques: Immunofluorescence, Staining, Cell Culture, Positive Control, Fluorescence, Western Blot, Control

Journal: bioRxiv

Article Title: Small-molecule IGF1R inhibitors extend healthspan in a mouse model

doi: 10.64898/2026.04.13.718278

Figure Lengend Snippet: Dose-response curves for Vegfa expression in adipose tissue (A and B) and free serum Igf-1 (C and D) in response to dietary exposure to different doses of NVP-ADW742 (A and C) and PPP (B and D). Each datapoint on the graphs is mean ± SE (n=5/group). Red datapoints are significantly different from controls (T-test p ≤ 0.05).

Article Snippet: Free blood IGF-1 was measured in murine serum using RayBio® Mouse IGF-1 ELISA Kit (Catalog #: ELM-IGF1).

Techniques: Expressing

Journal: bioRxiv

Article Title: Midazolam suppresses glioma progression by attenuating neuronal activity and downregulating IGF1 signaling

doi: 10.64898/2026.03.31.715727

Figure Lengend Snippet: (A) Differentially expressed genes and overall expression trends of cortical neurons from seven different groups. “D vs A”: log₂FC of KCls vs Control, “G vs D”: log₂FC of MDZs vs KCls. (B) Gene expression profiles of commonly secreted neuronal factors under KCl, and MDZ conditions. IGF1 is highlighted by a red dashed line. (C) Validation of IGF1and BDNF expression by RT-qPCR. n.s. P >0.05, * P < 0.05, ** P < 0.01, **** P < 0.0001. n=3. (D-E) Representative images and quantification of EdU assay after IGFBP3 experiment in GL261 cells. Scale bar = 100 μm. (F-G) Representative images and quantification of EdU assay after Linsitinib (OSI-906) treated experiments in GL261 cells. Scale bar = 200 μm. (H-I) Activation of the IGF1 downstream PI3K/AKT signaling pathway in glioma cells and its quantitative analysis. (J-K) PI3K/AKT signaling pathway change after IGFBP3 incubation in glioma cells. n.s. P >0.05, * P< 0.05, ** P< 0.01, *** P< 0.001, **** P< 0.0001. n=3.

Article Snippet: The concentrations of IGF1 in Neu-CMs were quantified using a commercial Mouse/Rat IGF1 ELISA Kit (Multi Sciences, China) according to the manufacturer’s instructions.

Techniques: Expressing, Control, Gene Expression, Biomarker Discovery, Quantitative RT-PCR, EdU Assay, Activation Assay, Incubation

Journal: bioRxiv

Article Title: Midazolam suppresses glioma progression by attenuating neuronal activity and downregulating IGF1 signaling

doi: 10.64898/2026.03.31.715727

Figure Lengend Snippet: (A) Overall distribution of IGF1 - related genes in all samples. (B) Standardized expression profile and function annotation of IGF1co-expression genes. (C-D) Over-representation enrichment analysis of genes of interest using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases in KCl treated group and MDZ treated group, respectively. (E) Fuzzy cluster analysis identifying five distinct expression patterns of genes participated in MAPK signaling pathway. (F) Transcriptional activity of Fos was evaluated using a univariable linear model. (G) Predicted c-Fos binding motif within the Igf1 promoter region identified using the JASPAR database. (H) ChIP-qPCR analysis assessing c-Fos enrichment at the predicted regulatory region. * P <0.05, ** P < 0.01, **** P< 0.0001. n=3.

Article Snippet: The concentrations of IGF1 in Neu-CMs were quantified using a commercial Mouse/Rat IGF1 ELISA Kit (Multi Sciences, China) according to the manufacturer’s instructions.

Techniques: Expressing, Activity Assay, Binding Assay, ChIP-qPCR

Journal: Development (Cambridge, England)

Article Title: Kupffer cells control neonatal hepatic metabolism via Igf1 signaling

doi: 10.1242/dev.204962

Figure Lengend Snippet: KC-hepatocyte crosstalk is altered in P0 KO Spi1 livers. (A) Bar plot showing the relative information flow of between WT Spi1 and KO Spi1 of inferred cell–cell communication using CellChat. (B) Comparison of the significant ligand–receptor pairs between WT Spi1 and KO Spi1 , which contribute to the signaling from KCs to the hepatocyte clusters. (C) Heatmap showing the relative importance of KC and hepatocyte clusters as sender, receiver, mediator and influencer, based on the computed four network centrality measures of IGF (top) and visfatin (bottom) signaling. (D) Box plot of variance stabilizing transformation-normalized Igf1 expression in hepatocytes and macrophages in WT Spi1 and KO Spi1 mice at P0. n =5 per genotype from 3 independent litters. Differential expression was tested using DESeq2 on raw counts. The whiskers represent the 5-95% percentile, the box extends from the 25th to 75th percentiles and the horizontal line represents the median. (E) Serum insulin levels measured by ELISA on WT Spi1 and KO Spi1 at P0. n =4-5 per genotype from 4 independent litters. Bar plot presented as mean±s.d. Unpaired Student's t -test. (F) Serum glucagon levels measured by ELISA on WT Spi1 and KO Spi1 at P0. n =6 per genotype from 3 independent litters. Bar plot presented as mean±s.d. Unpaired Student's t -test. (G) Enrichment analysis of downregulated phosphorylation sites showing the decreased and increased phosphorylation in KO Spi1 liver compared to WT Spi1 . n =4-6 per genotype from 5 independent litters.

Article Snippet: To determine the amount of Igf1 present in the perinatal liver and serum the Quantitative ELISA Mouse/Rat Igf1 Kit Liver Glycogen Assay Kit from R&D Systems (MG100) was used.

Techniques: Comparison, Transformation Assay, Expressing, Quantitative Proteomics, Enzyme-linked Immunosorbent Assay, Phospho-proteomics

Journal: Development (Cambridge, England)

Article Title: Kupffer cells control neonatal hepatic metabolism via Igf1 signaling

doi: 10.1242/dev.204962

Figure Lengend Snippet: KC-derived Igf1 regulates glycogen homeostasis in hepatocytes at birth. (A) Percentage of (left) and normalized (right) Igf1 expression in the respective hepatic cell type during embryogenesis. (B) Breeding scheme to produce KO Igf1 mice and littermate controls ( WT Igf1 ). Created in BioRender by Mass, E., 2025. https://BioRender.com/jvsfc8p . This figure was sublicensed under CC-BY 4.0 terms. (C,D) Igf1 levels measured by ELISA on whole liver lysate (C) and serum (D) of WT Igf1 and KO Igf1 at P0. n =7-8 per genotype from 4 independent litters. Bar plots presented as mean±s.d. Unpaired Student's t -test. (E) Glycogen levels measured on whole liver lysates of WT Igf1 and KO Igf1 at P0. n =11-16 per genotype from 7 independent litters. Values were normalized per litter. The whiskers represent the 5-95% percentile, the box extends from the 25th to 75th percentiles and the horizontal line represents the median. Cross indicates the mean. Mann–Whitney test. (F) Representative transmission electron micrograph from WT Igf1 and KO Igf1 livers at P0. n =3-4 per genotype from 2 independent litters. GP, glycogen particle; N, nucleus. Scale bars: 8 µm. (G) Scheme indicating the quantification process of glycogen content in hepatocytes. (H) Hepatocyte glycogen content of KO Igf1 normalized to WT Igf1 littermates; each value represents one hepatocyte (ten hepatocytes were assessed per liver). n =3-4 per genotype from 2 independent litters. Mann–Whitney test. (I) Normalized total metabolite abundance in WT Igf1 and KO Igf1 livers following [U- 13 C 6 ]-glucose tracing at P0. n =5-6 per genotype from 2 independent litters. Unpaired Student's t -test. ns, not significant ( P >0.05). (J) Fractional enrichment of labeled metabolites following [U- 13 C 6 ]-glucose tracing at P0 with and without the addition of exogenous Igf1 protein. Liver samples with and without Igf1 from the same animal are connected with a line. n =5-6 per genotype from 2 independent litters. Wilcoxon test.

Article Snippet: To determine the amount of Igf1 present in the perinatal liver and serum the Quantitative ELISA Mouse/Rat Igf1 Kit Liver Glycogen Assay Kit from R&D Systems (MG100) was used.

Techniques: Derivative Assay, Expressing, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Transmission Assay, Labeling