Journal: Respiratory Research

Article Title: ROBO2 signaling in lung development regulates SOX2/SOX9 balance, branching morphogenesis and is dysregulated in nitrofen-induced congenital diaphragmatic hernia

doi: 10.1186/s12931-020-01568-w

Figure Lengend Snippet: Effect of ROBO1 or ROBO2 functional impairment in branching morphogenesis. a The upper panel is representative of untreated lung explants (0 ng/mL) at D0; the bottom panel represents lung explants treated with recombinant IgG protein (0 ng/mL) and several doses of recombinant ROBO1 or ROBO2 proteins at day 4 (D4). b Morphometric analysis of the number of peripheral airway buds of fetal rat lung explants treated with increasing concentrations of recombinant ROBO1 (black) and ROBO2 (gray) proteins. Results are expressed as the D4/D0 ratio. c Examples of representative blots are showed for each analysed protein. Protein expression levels of d ROBO; e SOX2; f SOX9; g total β-Catenin; h non-phospho (active) β-Catenin; and i BMP4 in normal explant cultures treated with recombinant rat ROBO1 Fc Chimera for ROBO1 inhibition (black line) or recombinant human ROBO2 Fc Chimera for ROBO2 functional impairment (gray line). Recombinant human IgG Fc was used as control (IgG). n ≥ 9 per protein/condition. Each lane represents a pooled-tissue sample, and relative expression was determined against β-Tubulin and IgG. The data are presented as means ± SEM. p < 0.05: α vs. IgG

Article Snippet: Blots were blocked in 5% bovine serum albumin and probed with primary antibodies to ROBO1 (1:500, ON, 4 oC; Cat No. sc25672, Santa Cruz Biotechnology Inc., USA), ROBO2 (1:250, ON, 4 oC; Cat No. sc16615, Santa Cruz Biotechnology Inc. USA), SOX2 (1:250, ON, 4 oC; Cat No. AF2018, R&D system, USA), SOX9 (1:250, ON, 4 oC; Cat No. AF3075, R&D system, USA), non-phospho (Active) β-Catenin (Ser33/37/Thr41) (1∶5000; Cat No. #4270, Cell Signaling Technology Inc., USA), total β-Catenin (1∶30,000; Cat No. #NBP1-54,467, NOVUS Biologicals, USA), and BMP4 (1:250, ON, 4 oC; Cat No. sc-6896, Santa Cruz Biotechnology Inc., USA) according to the manufacturer's instructions.

Techniques: Functional Assay, Recombinant, Expressing, Inhibition, Control

Journal: Respiratory Research

Article Title: ROBO2 signaling in lung development regulates SOX2/SOX9 balance, branching morphogenesis and is dysregulated in nitrofen-induced congenital diaphragmatic hernia

doi: 10.1186/s12931-020-01568-w

Figure Lengend Snippet: a Overview of the main changes in spatiotemporal distribution of ROBO1, ROBO2, SOX2 and SOX9 at pseudoglandular (E17.5) and saccular stages (E21.5) in hypoplastic (hyp) fetal lungs (lower panel). w/o without b A proposed model of ROBO regulation of ex vivo branching morphogenesis through BMP4, β-Catenin, SOX2 and SOX9

Article Snippet: Blots were blocked in 5% bovine serum albumin and probed with primary antibodies to ROBO1 (1:500, ON, 4 oC; Cat No. sc25672, Santa Cruz Biotechnology Inc., USA), ROBO2 (1:250, ON, 4 oC; Cat No. sc16615, Santa Cruz Biotechnology Inc. USA), SOX2 (1:250, ON, 4 oC; Cat No. AF2018, R&D system, USA), SOX9 (1:250, ON, 4 oC; Cat No. AF3075, R&D system, USA), non-phospho (Active) β-Catenin (Ser33/37/Thr41) (1∶5000; Cat No. #4270, Cell Signaling Technology Inc., USA), total β-Catenin (1∶30,000; Cat No. #NBP1-54,467, NOVUS Biologicals, USA), and BMP4 (1:250, ON, 4 oC; Cat No. sc-6896, Santa Cruz Biotechnology Inc., USA) according to the manufacturer's instructions.

Techniques: Ex Vivo

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

Journal: International Journal of Molecular Sciences

Article Title: Tankyrase Regulates Neurite Outgrowth through Poly(ADP-ribosyl)ation-Dependent Activation of β-Catenin Signaling

doi: 10.3390/ijms23052834

Figure Lengend Snippet: β-catenin pathway mediates neurite outgrowth in PC12 cells. ( A ) Effect of β-catenin inhibitors on TNKS1/2 localization and neurite outgrowth in PC12 cells. Cells were treated with ICG-001 (5 μM) and PNU-74654 (20 μM) after 1 day in culture with NGF (100 ng/mL). After 4 days, cells were stained with anti-TNKS1/2 and anti-MAP2 antibodies. Nuclei were stained with DAPI. Scale bar: 20 μm. ( B ) Mean neurite length in PC12 cells. Data are shown as means ± SEM ( n = 14–18). *** p < 0.001 vs. Control (one-way ANOVA with post hoc Dunnett’s test).

Article Snippet: PJ34 was purchased from Enzo Life Sciences (Farmingdale, NY, USA); ABT888 from APExBIO (Houston, TX, USA); XAV939 from Cayman chemical company (Ann Arbor, MI, USA); MG-132, ICG-001, and PNU-74654 from Selleckchem (Houston, TX, USA); nerve growth factor-7S (NGF) from Sigma-Aldrich (Burlington, MA, USA); rabbit polyclonal anti-TNKS1/2 antibody (H-350), mouse monoclonal anti-MAP2 antibody (AP20), mouse monoclonal anti-PAR antibody (10H), and ADP-HPD from Santa Cruz Biotechnology (Dallas, TX, USA); rabbit monoclonal anti-Axin1 antibody (C76H11), and rabbit monoclonal anti-unphosphorylated (Ser33/Thr41) β-catenin antibody (D13A1) from Cell Signaling Technology (Danvers, MA); DAPI, Alexa Fluor 488-conjugated goat anti-rabbit IgG, and Alexa Fluor 568-conjugated goat anti-mouse IgG from Thermo Fisher Scientific (Waltham, MA, USA)); and predesigned primers for real-time PCR from Takara Bio (Kusatsu, Japan).

Techniques: Staining, Control

Journal: International Journal of Molecular Sciences

Article Title: Tankyrase Regulates Neurite Outgrowth through Poly(ADP-ribosyl)ation-Dependent Activation of β-Catenin Signaling

doi: 10.3390/ijms23052834

Figure Lengend Snippet: TNKS1/2 regulate β-catenin signaling through poly(ADP-ribosyl)ation in PC12 cells. ( A ) Effect of XAV939 on protein expression of Axin1 and unphosphorylated (non- p )-β-catenin in PC12 cells. Cells were treated with XAV939 (10 μM) for 1 day, followed by Western blotting with the indicated antibodies. GAPDH was used as a loading control. ( B ) Relative protein expression of Axin1 and non-p-β-catenin. Data are shown as means ± SEM ( n = 3). ** p < 0.01 vs. Control (Student’s t -test). ( C ) Effect of XAV939 on subcellular localization of non-p-β-catenin in PC12 cells. Cells were treated for 1 day with XAV939 (10 μM) and then stained using an anti-non-p-β-catenin antibody. Nuclei were stained with DAPI. Scale bar: 20 μm. ( D ) and ( F ) Effect of XAV939 on poly(ADP-ribosyl)ation of TNKS1/2 ( D ) and Axin1 ( F ) in PC12 cells. Cells were treated for 4 h with XAV939 (10 μM) and then subjected to macrodomain (MacroD)-GST pulldown assays followed by Western blotting with the indicated antibodies. ( E ) and ( G ) Relative poly(ADP-ribosyl)ation levels on TNKS1/2 ( E ) and Axin1 ( G ). Data are shown as means ± SEM ( n = 4). * p < 0.05, *** p < 0.001 vs. Control (Student’s t -test). ( H ) Effect of PARP inhibitors on NrCAM mRNA expression in PC12 cells. Cells were treated for 24 h with the indicated PARP inhibitors (10 μM) and then subjected to real-time PCR using NrCAM primers. NrCAM mRNA levels were normalized to GAPDH mRNA. Data are shown as means ± SEM ( n = 3) * p < 0.05, *** p < 0.001 vs. Control (one-way ANOVA with post hoc Dunnett’s test).

Article Snippet: PJ34 was purchased from Enzo Life Sciences (Farmingdale, NY, USA); ABT888 from APExBIO (Houston, TX, USA); XAV939 from Cayman chemical company (Ann Arbor, MI, USA); MG-132, ICG-001, and PNU-74654 from Selleckchem (Houston, TX, USA); nerve growth factor-7S (NGF) from Sigma-Aldrich (Burlington, MA, USA); rabbit polyclonal anti-TNKS1/2 antibody (H-350), mouse monoclonal anti-MAP2 antibody (AP20), mouse monoclonal anti-PAR antibody (10H), and ADP-HPD from Santa Cruz Biotechnology (Dallas, TX, USA); rabbit monoclonal anti-Axin1 antibody (C76H11), and rabbit monoclonal anti-unphosphorylated (Ser33/Thr41) β-catenin antibody (D13A1) from Cell Signaling Technology (Danvers, MA); DAPI, Alexa Fluor 488-conjugated goat anti-rabbit IgG, and Alexa Fluor 568-conjugated goat anti-mouse IgG from Thermo Fisher Scientific (Waltham, MA, USA)); and predesigned primers for real-time PCR from Takara Bio (Kusatsu, Japan).

Techniques: Expressing, Western Blot, Control, Staining, Real-time Polymerase Chain Reaction

Journal: International Journal of Molecular Sciences

Article Title: Tankyrase Regulates Neurite Outgrowth through Poly(ADP-ribosyl)ation-Dependent Activation of β-Catenin Signaling

doi: 10.3390/ijms23052834

Figure Lengend Snippet: Tankyrase activates β-catenin signaling to enhance neurite outgrowth and synapse formation. TNKS1/2 catalyze poly(ADP-ribosyl)ation of Axin and TNKS1/2 themselves, which activates β catenin signaling. Once translocated to the nucleus, β-catenin promotes transcription of NrCAM gene. β-catenin is also involved in stabilizing cadherin on the cell membrane. TNKS1/2-mediated upregulation of adhesion molecules may enhance neurite outgrowth and synapse formation.

Article Snippet: PJ34 was purchased from Enzo Life Sciences (Farmingdale, NY, USA); ABT888 from APExBIO (Houston, TX, USA); XAV939 from Cayman chemical company (Ann Arbor, MI, USA); MG-132, ICG-001, and PNU-74654 from Selleckchem (Houston, TX, USA); nerve growth factor-7S (NGF) from Sigma-Aldrich (Burlington, MA, USA); rabbit polyclonal anti-TNKS1/2 antibody (H-350), mouse monoclonal anti-MAP2 antibody (AP20), mouse monoclonal anti-PAR antibody (10H), and ADP-HPD from Santa Cruz Biotechnology (Dallas, TX, USA); rabbit monoclonal anti-Axin1 antibody (C76H11), and rabbit monoclonal anti-unphosphorylated (Ser33/Thr41) β-catenin antibody (D13A1) from Cell Signaling Technology (Danvers, MA); DAPI, Alexa Fluor 488-conjugated goat anti-rabbit IgG, and Alexa Fluor 568-conjugated goat anti-mouse IgG from Thermo Fisher Scientific (Waltham, MA, USA)); and predesigned primers for real-time PCR from Takara Bio (Kusatsu, Japan).

Techniques: Membrane

Journal: eLife

Article Title: T-ALL leukemia stem cell 'stemness' is epigenetically controlled by the master regulator SPI1

doi: 10.7554/eLife.38314

Figure Lengend Snippet: The biological properties of the newly defined HAVCR2 high LSC subgroup in comparison to other subgroups in the Pten -null T-ALL model.

Article Snippet: Antibody , Nonphospho (active)-β- catenin , Cell Signaling Technology , 70034 s , .

Techniques: Comparison, Marker, Activity Assay

Journal: eLife

Article Title: T-ALL leukemia stem cell 'stemness' is epigenetically controlled by the master regulator SPI1

doi: 10.7554/eLife.38314

Figure Lengend Snippet: ( A ) q-PCR analysis of SPI1 and SPI1-regulated HAVCR2 and c-MYC expressions after the overexpression of active β-catenin in the Jurkat T-ALL cell line (red bars). The data are normalized to that of empty plasmid controls (blue bars); ( B–E ) Upper panels: quantitative intracellular FACS analyses of P-GSK-3β, non-phospho-β-catenin, P-p65 and SPI1 levels in the HAVCR2 high , HAVCR2 low and blast subgroups; lower panels: representative intracellular FACS analysis of P-GSK-3β, non-phospho-β-catenin, P-p65 and SPI1 levels in the HAVCR2 high , HAVCR2 low and blast subgroups. Gray line, isotype control;( F ) FACS analysis shows cells in the HAVCR2 high subgroup at the ETP/DN1 stage, which are absent in WT and dKO mice; ( G ) Representative FACS plots show the number of cells in the HAVCR2 high subgroup in the different drug treatment groups. The data in A, B, C, D and E are the means ± S.Ds of 3 independent tests; *p≤0.05; **p≤0.01; ***p≤0.001.

Article Snippet: Antibody , Nonphospho (active)-β- catenin , Cell Signaling Technology , 70034 s , .

Techniques: Over Expression, Plasmid Preparation, Control

Journal: eLife

Article Title: T-ALL leukemia stem cell 'stemness' is epigenetically controlled by the master regulator SPI1

doi: 10.7554/eLife.38314

Figure Lengend Snippet: ( A ) Comparison of the HAVCR2 high subgroup population (left panel) and of the levels of non-phosphorylated β -catenin (middle panel) and SPI1 (right panel) within the HAVCR2 high subgroup without (upper panels) and with BAY6060 treatment (low panels); ( B ) Survival curve for Cdh5-Cre + ;Pten L/L mice treated with BAY6060 and BAY1082439 alone and in combination; ( C ) Survival curve for Cdh5-Cre + ;Pten L/L mice treated with rapamycin in combination with either an IgG control antibody or an anti-HAVCR2 antibody.

Article Snippet: Antibody , Nonphospho (active)-β- catenin , Cell Signaling Technology , 70034 s , .

Techniques: Comparison, Control

Journal: eLife

Article Title: T-ALL leukemia stem cell 'stemness' is epigenetically controlled by the master regulator SPI1

doi: 10.7554/eLife.38314

Figure Lengend Snippet:

Article Snippet: Antibody , Nonphospho (active)-β- catenin , Cell Signaling Technology , 70034 s , .

Techniques: