Journal: iScience

Article Title: Panx1 deficiency exacerbates GAN diet-induced obesity by destabilizing β-catenin via GSK3β

doi: 10.1016/j.isci.2026.115098

Figure Lengend Snippet: Panx1 knockout downregulated the Wnt/β-Catenin pathway in eWAT tissue (A) PCA of gene expression profiles ( n = 6/group). (B) GSEA enrichment of the Wnt/β-catenin pathway in eWAT. (C) Venn diagram showing overlap of DEGs with Wnt/β-catenin pathway genes. (D) Heatmap of Wnt-related DEGs in WT and Panx1 −/− mice. (E) Western blot of active β-catenin and downstream proteins (β-actin as loading control). Data are mean ± SD, statistics in (E): ∗ p < 0.05, ∗∗ p < 0.01, two-sided Student’s t test.

Article Snippet: Non-phospho (Active) β-Catenin (Ser33/37/Thr41) Rabbit mAb , Cell Signaling Technology , Cat# 8814S; RRID: AB_11127856.

Techniques: Knock-Out, Gene Expression, Western Blot, Control

Journal: iScience

Article Title: Panx1 deficiency exacerbates GAN diet-induced obesity by destabilizing β-catenin via GSK3β

doi: 10.1016/j.isci.2026.115098

Figure Lengend Snippet: Panx1 increases β-Catenin activity and promotes its nuclear translocation (A) Western blot shows Panx1 overexpression and knockout efficiency (GAPDH as loading control). (B) Glycosylation patterns of exogenous Panx1. (C and D) Total and non-phosphorylated β-catenin levels in whole cell and nuclear fractions (Lamin B1 as loading control). (E) Immunofluorescence of β-catenin nuclear translocation (red; nuclei, blue). Scale bars, 25 μm. (F) qRT-PCR of Wnt/β-catenin downstream gene expression normalized to PPIA. (G) Western blot of Wnt/β-catenin downstream proteins (GAPDH as loading control). Data are mean ± SD, statistics in (C), (D), (F), and (G): ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001, two-sided Student’s t test.

Article Snippet: Non-phospho (Active) β-Catenin (Ser33/37/Thr41) Rabbit mAb , Cell Signaling Technology , Cat# 8814S; RRID: AB_11127856.

Techniques: Activity Assay, Translocation Assay, Western Blot, Over Expression, Knock-Out, Control, Glycoproteomics, Immunofluorescence, Quantitative RT-PCR, Gene Expression

Journal: iScience

Article Title: Panx1 deficiency exacerbates GAN diet-induced obesity by destabilizing β-catenin via GSK3β

doi: 10.1016/j.isci.2026.115098

Figure Lengend Snippet: Panx1 forms complexes with β-Catenin and GSK-3β (A) Western blot of destruction complex proteins in Panx1-OE and Panx1 −/− 3T3-L1 cells (GAPDH as loading control). (B) Co-IP of Panx1 with β-catenin or destruction complex proteins in HEK293T cells expressing Panx1-HA. (C) Co-IP of β-catenin (total or phosphorylated) with GSK-3β in Panx1-OE and Panx1 −/− cells (GAPDH as loading control for input). (D) Western blot showing β-catenin knockdown efficiency in Panx1-OE cells (GAPDH as loading control). (E) Co-IP of Panx1 with GSK-3β after β-catenin knockdown (GAPDH as loading control for input). (F) Predicted 3D structures of Panx1, β-catenin, and GSK-3β complexes showing conformational changes. (G) Ranking scores of binding affinities among Panx1, β-catenin, and GSK-3β. Data are mean ± SD, statistics in (A): ∗ p < 0.05 and ∗∗ p < 0.01, two-sided Student’s t test.

Article Snippet: Non-phospho (Active) β-Catenin (Ser33/37/Thr41) Rabbit mAb , Cell Signaling Technology , Cat# 8814S; RRID: AB_11127856.

Techniques: Western Blot, Control, Co-Immunoprecipitation Assay, Expressing, Knockdown, Binding Assay

Journal: iScience

Article Title: Panx1 deficiency exacerbates GAN diet-induced obesity by destabilizing β-catenin via GSK3β

doi: 10.1016/j.isci.2026.115098

Figure Lengend Snippet: Effect of Panx1 on cell proliferation (A) Cell viability of Panx1-OE and Panx1 −/− cells. (B–D) Flow cytometry analysis of cell cycle distribution (PI staining) with the quantification of G1, S, and G2/M phases. (E) Western blot confirms β-catenin knockdown and effects on downstream proteins in Panx1-OE cells. (F and G) Flow cytometry and quantification of cell cycle distribution after β-catenin knockdown. Each panel represents an independently generated pair of stable lines, and statistical analyses are performed within each matched pair. Data are mean ± SD, statistics in (A), (D), and (G): ∗∗ p < 0.01 and ∗∗∗ p < 0.001, two-sided Student’s t test.

Article Snippet: Non-phospho (Active) β-Catenin (Ser33/37/Thr41) Rabbit mAb , Cell Signaling Technology , Cat# 8814S; RRID: AB_11127856.

Techniques: Flow Cytometry, Staining, Western Blot, Knockdown, Generated

Journal: Inflammation Research

Article Title: Macrophage β-catenin-Ihh axis induces hepatic stellate cell activation and fibrosis in metabolic dysfunction-associated steatohepatitis

doi: 10.1007/s00011-026-02220-x

Figure Lengend Snippet: Macrophage β-catenin is activated in HFD-induced MASH. Wild-type mice were provided with either a normal chow diet (NCD) or a high-fat diet (HFD) for 28 weeks. A Double immunofluorescence staining was performed on liver sections. Antibodies against F4/80 (displayed in green) and A-β-catenin (shown in red) were used. The cell nuclei were stained with DAPI (appearing blue). Five mice were included in each group. The scale bar represents 200 μm. B Liver macrophages were separated from mice that had been fed NCD or HFD. The mRNA level of Ctnnb1 (the gene encoding β-catenin) in liver macrophages of wild-type mice that had received NCD or HFD for 28 weeks was measured ( n = 5 samples/group). C Western blot analysis was carried out to detect the expression of A-β-catenin in liver macrophages from wild-type mice fed with NCD or HFD ( n = 5 samples/group). The data are presented as the mean ± standard deviation (SD). ** P < 0.01

Article Snippet: By using rat monoclonal antibody for F4/80 (Cat#: sc-52664, dilution 1:50, Santa Cruz Biotechnology, California) as well as rabbit monoclonal antibody for active β-catenin (non-phosphorylated serine 33/37/threonine41) (Cat#: 8814, 1:200 dilution, Cell Signaling Technology, Massachusetts) or anti-Ihh antibody (Cat#: ab39634, 1:100 dilution, Abcam, Massachusetts), the double-positive macrophages for F4/80 and A-β-catenin or Ihh were successfully identified.

Techniques: Double Immunofluorescence Staining, Staining, Western Blot, Expressing, Standard Deviation

Journal: Inflammation Research

Article Title: Macrophage β-catenin-Ihh axis induces hepatic stellate cell activation and fibrosis in metabolic dysfunction-associated steatohepatitis

doi: 10.1007/s00011-026-02220-x

Figure Lengend Snippet: Deficiency of myeloid β-catenin alleviates hepatic steatosis induced by HFD. A Serum ALT and AST levels were measured in β-catenin FL/FL and β-catenin M−KO mice following 28 weeks of normal chow diet (NCD) or high-fat diet (HFD) feeding ( n = 5 samples/group). B Liver-to-body weight ratios were examined in β-catenin FL/FL and β-catenin M−KO mice fed with NCD or HFD ( n = 5 samples/group). C Liver triglyceride (TG) and total cholesterol (TC) levels were determined in β-catenin FL/FL and β-catenin M−KO mice fed with NCD or HFD ( n = 5 samples/group). D Representative hematoxylin and eosin (H&E) staining of liver sections and the non-alcoholic fatty liver disease activity score (NAS) were obtained from β-catenin FL/FL and β-catenin M−KO mice fed with NCD or HFD ( n = 5 samples/group). The scale bars represent 100 μm. E Representative Oil Red O-stained images of liver sections were taken from β-catenin FL/FL and β-catenin M−KO mice fed with NCD or HFD ( n = 5 samples/group). The scale bar is 50 μm. The data are presented as mean ± standard deviation (SD). ** P < 0.01

Article Snippet: By using rat monoclonal antibody for F4/80 (Cat#: sc-52664, dilution 1:50, Santa Cruz Biotechnology, California) as well as rabbit monoclonal antibody for active β-catenin (non-phosphorylated serine 33/37/threonine41) (Cat#: 8814, 1:200 dilution, Cell Signaling Technology, Massachusetts) or anti-Ihh antibody (Cat#: ab39634, 1:100 dilution, Abcam, Massachusetts), the double-positive macrophages for F4/80 and A-β-catenin or Ihh were successfully identified.

Techniques: Staining, Activity Assay, Standard Deviation

Journal: Inflammation Research

Article Title: Macrophage β-catenin-Ihh axis induces hepatic stellate cell activation and fibrosis in metabolic dysfunction-associated steatohepatitis

doi: 10.1007/s00011-026-02220-x

Figure Lengend Snippet: Deficiency of myeloid β-catenin ameliorates MASH-associated liver fibrosis. A Representative Sirius Red staining of liver sections was shown from β-catenin FL/FL and β-catenin M−KO mice. These mice were fed with either a normal chow diet (NCD) or a high-fat diet (HFD) for 28 weeks ( n = 5 samples/group). Scale bars were 100 μm. B Immunofluorescence images of α-SMA + myofibroblasts in liver sections were obtained from NCD-fed or HFD-fed β-catenin FL/FL and β-catenin M−KO mice ( n = 5 samples/group). Scale bars measured 50 μm. C Relative mRNA levels of fibrogenic genes such as Acta2 , Col1α1 , Col3α 1, and Timp 1 were detected in the livers of β-catenin FL/FL and β-catenin M−KO mice that were fed with NCD or HFD ( n = 5 samples/group). Data were expressed as the mean ± standard deviation (SD). ** P < 0.01

Article Snippet: By using rat monoclonal antibody for F4/80 (Cat#: sc-52664, dilution 1:50, Santa Cruz Biotechnology, California) as well as rabbit monoclonal antibody for active β-catenin (non-phosphorylated serine 33/37/threonine41) (Cat#: 8814, 1:200 dilution, Cell Signaling Technology, Massachusetts) or anti-Ihh antibody (Cat#: ab39634, 1:100 dilution, Abcam, Massachusetts), the double-positive macrophages for F4/80 and A-β-catenin or Ihh were successfully identified.

Techniques: Staining, Immunofluorescence, Standard Deviation

Journal: Inflammation Research

Article Title: Macrophage β-catenin-Ihh axis induces hepatic stellate cell activation and fibrosis in metabolic dysfunction-associated steatohepatitis

doi: 10.1007/s00011-026-02220-x

Figure Lengend Snippet: Deficiency of myeloid β-catenin inhibits Ihh expression in macrophages in HFD-induced MASH. A Relative mRNA levels of Shh and Ihh in liver macrophages isolated from HFD-fed β-catenin FL/FL and β-catenin M−KO mice were determined. B Western blot was performed to analyze the expression of Ihh in liver macrophages from HFD-fed β-catenin FL/FL and β-catenin M−KO mice. C Double immunofluorescence staining was carried out on liver sections using antibodies against F4/80 (green) and Ihh (red). The nuclei were stained with DAPI (blue). There were five mice in each group. The scale bar represents 200 μm. The data are shown as the mean ± standard deviation (SD). ** P < 0.01

Article Snippet: By using rat monoclonal antibody for F4/80 (Cat#: sc-52664, dilution 1:50, Santa Cruz Biotechnology, California) as well as rabbit monoclonal antibody for active β-catenin (non-phosphorylated serine 33/37/threonine41) (Cat#: 8814, 1:200 dilution, Cell Signaling Technology, Massachusetts) or anti-Ihh antibody (Cat#: ab39634, 1:100 dilution, Abcam, Massachusetts), the double-positive macrophages for F4/80 and A-β-catenin or Ihh were successfully identified.

Techniques: Expressing, Isolation, Western Blot, Double Immunofluorescence Staining, Staining, Standard Deviation

Journal: Inflammation Research

Article Title: Macrophage β-catenin-Ihh axis induces hepatic stellate cell activation and fibrosis in metabolic dysfunction-associated steatohepatitis

doi: 10.1007/s00011-026-02220-x

Figure Lengend Snippet: Ihh signaling is crucial for macrophage β-catenin-mediated hepatic fibrosis in HFD-induced MASH. To restore Ihh expression in macrophages, mice were administered the Ihh plasmid via polyethylenimine nanoparticles, namely in vivo jetPEI-Man. A Representative Sirius Red staining images of liver sections from HFD-fed β-catenin FL/FL and β-catenin M−KO mice. These mice were either treated or not treated with jetPEI-Man-Ihh (jetPEI-Ihh) or jetPEI-Man-GFP (jetPEI-GFP) ( n = 5 mice per group). Scale bars are 100 μm. B Immunofluorescence images of α-SMA + myofibroblasts in liver sections from HFD-fed β-catenin FL/FL and β-catenin M−KO mice. These mice were either treated or not treated with jetPEI-Ihh or jetPEI-GFP ( n = 5 mice per group). Scale bars measure 50 μm. C Relative mRNA levels of fibrogenic genes such as Acta2 , Col1α1 , Col3α1 , and Timp1 in the livers of HFD-fed β-catenin FL/FL and β-catenin M−KO mice. These mice were either treated or not treated with jetPEI-Ihh or jetPEI-GFP ( n = 5 mice per group). Data are expressed as the mean ± standard deviation (SD). * P < 0.05, ** P < 0.01

Article Snippet: By using rat monoclonal antibody for F4/80 (Cat#: sc-52664, dilution 1:50, Santa Cruz Biotechnology, California) as well as rabbit monoclonal antibody for active β-catenin (non-phosphorylated serine 33/37/threonine41) (Cat#: 8814, 1:200 dilution, Cell Signaling Technology, Massachusetts) or anti-Ihh antibody (Cat#: ab39634, 1:100 dilution, Abcam, Massachusetts), the double-positive macrophages for F4/80 and A-β-catenin or Ihh were successfully identified.

Techniques: Expressing, Plasmid Preparation, In Vivo, Staining, Immunofluorescence, Standard Deviation

Journal: Inflammation Research

Article Title: Macrophage β-catenin-Ihh axis induces hepatic stellate cell activation and fibrosis in metabolic dysfunction-associated steatohepatitis

doi: 10.1007/s00011-026-02220-x

Figure Lengend Snippet: Macrophage β-catenin induces Ihh expression and secretion. Bone marrow-derived macrophages (BMDMs) were treated with LPS (100 ng/ml) and PA (250 µM) or PBS for 12 h. A The mRNA level of Ctnnb1 in BMDMs following treatment with PA/LPS or PBS. B Western blot analysis was carried out to assess the expression of A-β-catenin in BMDMs after treatment with PA/LPS or PBS. C BMDMs underwent a ChIP-PCR assay using an anti-β-catenin or IgG antibody to detect the potential β-catenin binding site within the Ihh intron 1. D The mRNA level of Ihh in BMDMs isolated from β-catenin FL/FL and β-catenin M−KO mice. E The concentration of Ihh in the media of BMDMs isolated from β-catenin FL/FL and β-catenin M−KO mice was measured by ELISA. The data are presented as the mean ± standard deviation (SD). ** P < 0.01

Article Snippet: By using rat monoclonal antibody for F4/80 (Cat#: sc-52664, dilution 1:50, Santa Cruz Biotechnology, California) as well as rabbit monoclonal antibody for active β-catenin (non-phosphorylated serine 33/37/threonine41) (Cat#: 8814, 1:200 dilution, Cell Signaling Technology, Massachusetts) or anti-Ihh antibody (Cat#: ab39634, 1:100 dilution, Abcam, Massachusetts), the double-positive macrophages for F4/80 and A-β-catenin or Ihh were successfully identified.

Techniques: Expressing, Derivative Assay, Western Blot, Binding Assay, Isolation, Concentration Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Inflammation Research

Article Title: Macrophage β-catenin-Ihh axis induces hepatic stellate cell activation and fibrosis in metabolic dysfunction-associated steatohepatitis

doi: 10.1007/s00011-026-02220-x

Figure Lengend Snippet: Macrophage β-catenin-induced Ihh promotes HSC activation. A The schematic illustration of the co-culture model that utilizes bone marrow-derived macrophages (BMDMs) and primary murine hepatic stellate cells (HSCs). B The relative mRNA levels of fibrogenic genes such as Acta2 , Col1α1 , and Col3α1 in HSCs were measured after co-culturing with BMDMs from β-catenin FL/FL and β-catenin M−KO mice. C Immunofluorescence pictures of α-SMA expression in HSCs were obtained after co-culturing with BMDMs from β-catenin FL/FL and β-catenin M−KO mice. The scale bars represent 100 μm. D Immunofluorescence images of α-SMA expression in HSCs were captured after co-culturing with BMDMs transfected with the Lv-β-catenin plasmid or the control vector (Lv-GFP). The scale bars are 100 μm. E Immunofluorescence images of α-SMA expression in HSCs were taken after co-culturing with Lv-β-catenin-transfected BMDMs, either in the presence or absence of Ihh neutralizing antibody. The scale bars measure 100 μm. F The mRNA levels of Gli1 , Gli2 , and Gli3 in HSCs were determined after co-culturing with BMDMs from β-catenin FL/FL and β-catenin M−KO mice. The data are presented as the mean ± standard deviation (SD). * P < 0.05, ** P < 0.01

Article Snippet: By using rat monoclonal antibody for F4/80 (Cat#: sc-52664, dilution 1:50, Santa Cruz Biotechnology, California) as well as rabbit monoclonal antibody for active β-catenin (non-phosphorylated serine 33/37/threonine41) (Cat#: 8814, 1:200 dilution, Cell Signaling Technology, Massachusetts) or anti-Ihh antibody (Cat#: ab39634, 1:100 dilution, Abcam, Massachusetts), the double-positive macrophages for F4/80 and A-β-catenin or Ihh were successfully identified.

Techniques: Activation Assay, Co-Culture Assay, Derivative Assay, Immunofluorescence, Expressing, Transfection, Plasmid Preparation, Control, Standard Deviation

Journal: Inflammation Research

Article Title: Macrophage β-catenin-Ihh axis induces hepatic stellate cell activation and fibrosis in metabolic dysfunction-associated steatohepatitis

doi: 10.1007/s00011-026-02220-x

Figure Lengend Snippet: Schematic representation of the macrophage β-catenin-Ihh axis in modulating the activation of hepatic stellate cells and fibrosis in MASH. The consumption of a high-fat diet (HFD) led to the activation of β-catenin in liver macrophages. The activation of macrophage β-catenin triggered the expression and secretion of Ihh, which in turn facilitated the activation of hepatic stellate cells and fibrosis in MASH

Article Snippet: By using rat monoclonal antibody for F4/80 (Cat#: sc-52664, dilution 1:50, Santa Cruz Biotechnology, California) as well as rabbit monoclonal antibody for active β-catenin (non-phosphorylated serine 33/37/threonine41) (Cat#: 8814, 1:200 dilution, Cell Signaling Technology, Massachusetts) or anti-Ihh antibody (Cat#: ab39634, 1:100 dilution, Abcam, Massachusetts), the double-positive macrophages for F4/80 and A-β-catenin or Ihh were successfully identified.

Techniques: Activation Assay, Expressing