Journal: Cell Death & Disease

Article Title: Tubule-derived CCN1 drives renal repair via α v β 5 -STAT6-ARG1-dependent reprogramming of macrophages

doi: 10.1038/s41419-025-08340-2

Figure Lengend Snippet: A A diagram illustrating the setup of the experiment uxIRI. B Representative images of H&E staining, KIM1, F4/80, and CCN1 immunostaining in kidney tissues. C Renal CCN1 protein and mRNA expression. D Serum CCN1 levels in mice after I/R. n = 4 mice per group. E CCN1 expression in RTECs from AKI patients by single-cell RNA-seq. F Representative fluorescent micrographs and quantification analysis of CCN1 (yellow), AQP1 (red), and F4/80 (green) staining in I/R-AKI mice kidney. n = 4 mice per group. G CCN1 mRNA expression levels in HK-2 cells after H/R or H 2 O 2 stimulation. H CCN1 protein expression levels in HK-2 cells after H/R or H 2 O 2 stimulation. I Secreted CCN1 protein levels in culture supernatants of HK-2 cells after H/R or H 2 O 2 stimulation. n = 3 per group. Data are presented as mean ± SD.

Article Snippet: BMDMs were seeded in the top chamber and treated with 2 μg/mL CCN1 protein, 50 μM ARG1 inhibitor (Numidargistat, MedChemExpress), or a combination of both.

Techniques: Staining, Immunostaining, Expressing, RNA Sequencing

Journal: Cell Death & Disease

Article Title: Tubule-derived CCN1 drives renal repair via α v β 5 -STAT6-ARG1-dependent reprogramming of macrophages

doi: 10.1038/s41419-025-08340-2

Figure Lengend Snippet: A A diagram illustrating the CCN1 protein administration in mice after I/R. B Blood urea nitrogen and serum creatinine levels in mice after I/R. C Representative images and quantification analysis of H&E staining, and KIM1 immunostaining in kidney tissues. D Density ridge plots showing the expression distributions of core-enriched gene sets identified by GSEA. E Selected enrichment plots from GSEA based on the gene enrichment profiles on CCN1-treated I/R kidneys versus I/R controls. F Gating strategy for CD45 + F4/80 + CD11b + cells in the kidney after I/R by flow cytometry. G Subsequent gating for macrophage polarization markers, including CD206, Ly6C, CD86 and CCR2 in the kidney after I/R by flow cytometry. n = 4 mice per group. Data are presented as mean ± SD.

Article Snippet: BMDMs were seeded in the top chamber and treated with 2 μg/mL CCN1 protein, 50 μM ARG1 inhibitor (Numidargistat, MedChemExpress), or a combination of both.

Techniques: Staining, Immunostaining, Expressing, Flow Cytometry

Journal: Cell Death & Disease

Article Title: Tubule-derived CCN1 drives renal repair via α v β 5 -STAT6-ARG1-dependent reprogramming of macrophages

doi: 10.1038/s41419-025-08340-2

Figure Lengend Snippet: A A diagram of uxIRI setup in a tubule-specific CCN1 knockdown model induced by renal pelvic AAV9-Ksp-shCCN1 injection. B Blood urea nitrogen and serum creatinine levels in mice after I/R. Representative images and quantification analysis of C H&E staining, D KIM1 and E F4/80 immunostaining in kidney tissues. F Gating strategy for CD45 + F4/80 + CD11b + cells in the kidney after I/R by flow cytometry. G Subsequent gating for macrophage polarization markers, including CD86, CCR2, Ly6C, and CD206 in the kidney after I/R by flow cytometry. n = 4 mice per group. Data are presented as mean ± SD.

Article Snippet: BMDMs were seeded in the top chamber and treated with 2 μg/mL CCN1 protein, 50 μM ARG1 inhibitor (Numidargistat, MedChemExpress), or a combination of both.

Techniques: Knockdown, Injection, Staining, Immunostaining, Flow Cytometry

Journal: Cell Death & Disease

Article Title: Tubule-derived CCN1 drives renal repair via α v β 5 -STAT6-ARG1-dependent reprogramming of macrophages

doi: 10.1038/s41419-025-08340-2

Figure Lengend Snippet: A A diagram of BMDMs isolation and CCN1 protein treatment. B , C Flow cytometry analysis of Ly6C⁺MHCII⁻ monocytes and Ly6C⁻MHCII⁺ macrophages following CCN1 stimulation. n = 3 per group. D CCK-8 assay of cell viability and adherence after CCN1 treatment. n = 6 per group. E Contour plot of CD86⁺, CCR2⁺, Ly6C⁻ and CD206⁺ BMDMs by flow cytometry. F Histogram of CD86, CCR2, Ly6C and CD206 of BMDMs by flow cytometry. n = 3 per group. Data are presented as mean ± SD.

Article Snippet: BMDMs were seeded in the top chamber and treated with 2 μg/mL CCN1 protein, 50 μM ARG1 inhibitor (Numidargistat, MedChemExpress), or a combination of both.

Techniques: Isolation, Flow Cytometry, CCK-8 Assay

Journal: Cell Death & Disease

Article Title: Tubule-derived CCN1 drives renal repair via α v β 5 -STAT6-ARG1-dependent reprogramming of macrophages

doi: 10.1038/s41419-025-08340-2

Figure Lengend Snippet: A Volcano plot showing differentially expressed genes in BMDMs with or without CCN1 treatment under unstimulated or LPS and IFN-γ-stimulated conditions. B Heatmaps of marker genes of seven macrophage subpopulations for macrophage subpopulations. C Concentration of growth factors IGF-1 and PDGF-BB in BMDMs medium. D The mRNA expression of growth factors ( Igf1 , Pdgfb , Vegfa , Hgf , and Hbegf ) in BMDMs. E A diagram of the transwell co-culture system with CCN1-treated BMDMs in the top chamber and HK-2 cells in the bottom chamber. F Representative image of EdU immunofluorescence staining and quantification of EdU + proportion in HK-2 cells in the bottom chamber. G Flow cytometry analysis of EdU⁺ HK-2 cells in the co-culture system. n = 3 per group. H Representative images and quantitative analysis of Ki67 immunostaining in I/R mice kidney with AAV-shScr and AAV-shCCN1. I Representative images of Ki67 immunostaining in I/R mice kidney with CCN1 treatment and Clodronate Liposomes (CL). n = 4 mice per group. Data are presented as mean ± SD.

Article Snippet: BMDMs were seeded in the top chamber and treated with 2 μg/mL CCN1 protein, 50 μM ARG1 inhibitor (Numidargistat, MedChemExpress), or a combination of both.

Techniques: Marker, Concentration Assay, Expressing, Co-Culture Assay, Immunofluorescence, Staining, Flow Cytometry, Immunostaining, Liposomes

Journal: Cell Death & Disease

Article Title: Tubule-derived CCN1 drives renal repair via α v β 5 -STAT6-ARG1-dependent reprogramming of macrophages

doi: 10.1038/s41419-025-08340-2

Figure Lengend Snippet: A BayesPrism workflow of scRNA-seq ( GSE174324 ) and bulk RNA-seq transcriptomic data integration and deconvolution based on BayesPrism to infer joint. B Cell-type fraction of deconvolution analysis across all samples. C Comparison of cell-type fractions following CCN1 treatment under normal and LPS-stimulated conditions. n = 4 per group. * P < 0.05. D , E The protein and mRNA expression of ARG1 in BMDMs. F , G Contour plot and histogram of flow cytometry of the proportion of ARG1 + cells in BMDMs. H The concentration of urea and Arginase activity in BMDMs. n = 4 per group. Data are presented as mean ± SD. I Representative fluorescent micrographs and quantification analysis of ARG1 (yellow), F4/80 (red) and EGFP-tag staining in I/R mice kidney with AAV-shScr and AAV-shCCN1 injection. J Flow cytometry of I/R mice kidney with CCN1 treatment. n = 4 mice per group. Data are presented as mean ± SD.

Article Snippet: BMDMs were seeded in the top chamber and treated with 2 μg/mL CCN1 protein, 50 μM ARG1 inhibitor (Numidargistat, MedChemExpress), or a combination of both.

Techniques: RNA Sequencing, Comparison, Expressing, Flow Cytometry, Concentration Assay, Activity Assay, Staining, Injection

Journal: Cell Death & Disease

Article Title: Tubule-derived CCN1 drives renal repair via α v β 5 -STAT6-ARG1-dependent reprogramming of macrophages

doi: 10.1038/s41419-025-08340-2

Figure Lengend Snippet: A GSEA enrichment of STAT-related pathways in CCN1-treated BMDMs. B Protein levels of p-STAT6, STAT6 and ARG1 in BMDMs. C Co-IP and MS workflow for CCN1-interacting proteins. D Overlap of CCN1-interacting proteins identified by Co-IP/MS and BioGRID database. E Molecular docking of CCN1 with ITGAV and ITGB5. F Co-IP validation of CCN1 binding to ITGAV and ITGB5 in BMDMs. G Protein levels of p-STAT6, STAT6 and ARG1 after α v β 5 integrin inhibitor treatment. n = 3 per group. Data are presented as mean ± SD.

Article Snippet: BMDMs were seeded in the top chamber and treated with 2 μg/mL CCN1 protein, 50 μM ARG1 inhibitor (Numidargistat, MedChemExpress), or a combination of both.

Techniques: Co-Immunoprecipitation Assay, Biomarker Discovery, Binding Assay

Journal: Molecular Therapy Oncology

Article Title: Targeted nanoencapsulation of tunicamycin reduces toxicity while improving its therapeutic effectiveness in pancreatic cancer cells

doi: 10.1016/j.omton.2025.201047

Figure Lengend Snippet: Effect of TM on 3D co-culture as in vitro desmoplasia and CCN1 expression in PDAC cells (A) In vitro desmoplasia assay using co-culture of untreated or NP TM-treated Panc-1 cells and fibroblast cells as depicted in upper panel. Arrow indicates a desmoplasia-like structure. Scale bars, 50 μm. (B) 3D co-culture of Panc-1 and pancreatic Stellate cells in True Gel3D hydrogel for the detection of tumor-stroma interaction and F-actin expression. Scale bars, 50 μm. (C) Immuno-western blot analysis of CCN1 in untreated, free, and nanoencapsulated TM-treated Panc-1 cell extracts. Cells were treated for 48 h, and 50 μg total protein/sample was separated in SDS-Page gel. (D) The graphs illustrated the signaling status of CCN1 in different samples derived from five independent experiments of each group. Data represent mean ± SD. The p value was calculated using the student t test .

Article Snippet: Anti-rabbit IgG (# 70745) was purchased from Cell Signaling, USA; anti-mouse IgG (# sc-516102) from Santa Cruz, Inc, USA; CCN1 polyclonal antibody (#26689-1-AP) and Neuropilin-1 (NRP-1) from ProteinTech, USA; and pSTAT3 polyclonal antibody (#9145) and STAT3 polyclonal antibody, peIF4E and EIF4E antibodies, and PCNA rabbit polyclonal antibody (#13110T) from Cell Signaling, Inc., USA.

Techniques: Co-Culture Assay, In Vitro, Expressing, Western Blot, SDS Page, Derivative Assay

Journal: European Journal of Medical Research

Article Title: The S1PR1–CCN1 axis drives endothelial-to-mesenchymal transition and vascular instability in brain arteriovenous malformations

doi: 10.1186/s40001-025-03484-5

Figure Lengend Snippet: CCN1 is upregulated upon S1PR1 silencing and correlates with EndoMT-associated endothelial activation in human bAVMs. A Proteomic profiling identifies CCN1 as significantly upregulated following S1PR1 knockdown in HUVECs. B , C Validation of CCN1 upregulation at the mRNA (qRT-PCR) and protein (Western blot) levels in S1PR1 knockdown endothelial cells ( n = 3). D Quantification of CCN1 expression in ruptured versus unruptured human bAVM tissues ( n = 3). E Representative Immunohistochemistry images of CCN1 in human bAVM tissues, showing elevated expression in ruptured versus unruptured samples. F Immunofluorescence staining of human bAVM tissue shows co-localization of CCN1 with the endothelial marker CD31. G , H Re-analysis of publicly available single-cell RNA sequencing data comparing CCN1 expression across normal and bAVM vascular endothelium. I Schematic illustration of endothelial-specific ccn1 overexpression in zebrafish embryos using Tol2-mediated transgenesis. J Quantification of ccn1 mRNA levels in control and ccn1 -overexpressing embryos at 2 days post-fertilization (2 dpf) by qRT-PCR ( n = 3). K Light field images of zebrafish embryos at 2 dpf. Cranial hemorrhage (yellow arrow) is observed in the ccn1 -overexpressing group but not in controls. Right panel: Quantification of cerebral hemorrhage incidence ( n = 100). L Confocal images of cranial vasculature and red blood cells in Tg (gata1:Ds Red; flk1:EGFP) embryos at 2 dpf. Overexpression of ccn1 led to erythrocyte leakage and blood pooling in the cranial region. M Representative images of trunk vasculature in 5 dpf embryos. Overexpression of ccn1 caused significant dilation of intersomitic vessels (ISVs), as shown by increased vessel diameter. Right panel: Quantification of ISV diameter ( n = 6). N Confocal images of cerebral vasculature at 5 dpf showing increased cerebrovascular density in ccn1-overexpressing embryos. Right panel: Quantification of cerebrovascular fluorescence area ( n = 6). Scale bars: 100 μm

Article Snippet: Sections were blocked in 5% normal serum and incubated overnight at 4 °C with primary antibodies against S1PR1 or CCN1 (Proteintech, China).

Techniques: Activation Assay, Knockdown, Biomarker Discovery, Quantitative RT-PCR, Western Blot, Expressing, Immunohistochemistry, Immunofluorescence, Staining, Marker, RNA Sequencing, Over Expression, Control, Fluorescence

Journal: European Journal of Medical Research

Article Title: The S1PR1–CCN1 axis drives endothelial-to-mesenchymal transition and vascular instability in brain arteriovenous malformations

doi: 10.1186/s40001-025-03484-5

Figure Lengend Snippet: Rescue of S1PR1 deficiency-induced EndoMT and migration by CCN1 silencing. A – C mRNA and protein expression of S1PR1 and CCN1 in HUVECs following S1PR1 knockdown with or without co-transfection of sh-CCN1. D – F Rescue of EndoMT markers (N-cadherin, α-SMA, VE-cadherin) assessed by qRT-PCR ( n = 3). G Representative transwell migration images and quantification at 24 h and 48 h post-treatment ( n = 3). H Representative scratch wound images and quantification of wound closure at 0 h and 24 h ( n = 3)

Article Snippet: Sections were blocked in 5% normal serum and incubated overnight at 4 °C with primary antibodies against S1PR1 or CCN1 (Proteintech, China).

Techniques: Migration, Expressing, Knockdown, Cotransfection, Quantitative RT-PCR