Journal: PLOS ONE

Article Title: mGBP2 engages Galectin-9 for immunity against Toxoplasma gondii

doi: 10.1371/journal.pone.0316209

Figure Lengend Snippet: Primers.

Article Snippet: Additionally, WB analysis was performed with a polyclonal rabbit anti-mouse antibody against Gal9 (ARP54821_P050, Aviva Systems Biology).

Techniques: Sequencing, Construct

Journal: PLOS ONE

Article Title: mGBP2 engages Galectin-9 for immunity against Toxoplasma gondii

doi: 10.1371/journal.pone.0316209

Figure Lengend Snippet: mGBP2 -/- MEFs were reconstituted with GFP-mGBP2 WT or one of the indicated mGBP2 truncation mutants as well as one of the N-terminal mCherry fusion proteins mCherry-mGBP2 or mCherry-Gal9. 10 6 Cells were stimulated with IFN-γ for 16 h and infected with T . gondii ME49 for 4h, MOI 50. Cells were lysed and lysate supernatants were incubated o/n with either RFP-Trap ® beads (A, B) or with GFP-Trap ® beads (C-F) at 4°C. Pull-down (PD) samples and appropriate cell lysate supernatants were subjected to Western Blotting (WB). Blots were stained with α-GFP (A, D, F) or α-mCherry (B, C, E) antibodies. GTP-binding domain (GM), the middle domain and the C-terminal effector domain (ME) or the C-terminal effector domain only (E) (37).

Article Snippet: Additionally, WB analysis was performed with a polyclonal rabbit anti-mouse antibody against Gal9 (ARP54821_P050, Aviva Systems Biology).

Techniques: Infection, Incubation, Western Blot, Staining, Binding Assay

Journal: PLOS ONE

Article Title: mGBP2 engages Galectin-9 for immunity against Toxoplasma gondii

doi: 10.1371/journal.pone.0316209

Figure Lengend Snippet: gondii . Recruitment and colocalization of mGBP2 and Gal9 was analyzed after transduction of a GFP-mGBP2 fusion construct in mGBP2 -/- MEFs and additional transduction of mCherry-Gal9. MEFs were seeded and incubated on glass slides, stimulated with IFN-γ for 16 h and subsequently infected with T . gondii ME49 for 2 h. After fixation, infected cells were treated with an α-RFP V H H nanobody conjugated to eGFPBoosterAtto647N and with an α-GFP V H H nanobody conjugated to eGFPBoosterAtto488 for enhancement of the immunofluorescence of mCherry and GFP, respectively. Glass slides were analyzed by STED microscopy. Bars 2 μm. The graphs in the right panel depict a fluorescence intensity analysis of STED images on the far right with the ImageJ software (Fiji) for Atto488 and Atto647 fluorescence signals along the cross sections of PVMs as indicated. The colocalization thresholds were set to 7500-max for the Cherry and 5000-max for GFP in mCherry-Gal9 and GFP-mGBP2 expressing cells. Three representative examples from three independent experiments are shown.

Article Snippet: Additionally, WB analysis was performed with a polyclonal rabbit anti-mouse antibody against Gal9 (ARP54821_P050, Aviva Systems Biology).

Techniques: Transduction, Construct, Incubation, Infection, Immunofluorescence, Microscopy, Fluorescence, Software, Expressing

Journal: PLOS ONE

Article Title: mGBP2 engages Galectin-9 for immunity against Toxoplasma gondii

doi: 10.1371/journal.pone.0316209

Figure Lengend Snippet: gondii . (A) WT and NIH 3T3 cell line clones with verified CRISPR/Cas9 mediated inactivation of Gal9 (see main text and and Figs) were stimulated with IFN-γ for 16 h and subsequently infected with T . gondii ME49 MOI10 for 24 h. After fixation, T . gondii were stained with an α-SAGI antibody and the cell nuclei were labeled with DAPI. Glass slides were analyzed by confocal microscopy. The amounts of rosettes (replicative units) and single parasites inside the PV were determined in WT, Gal9- deficient NIH 3T3 fibroblasts 24 h after infection. In each cell line more than 100 PVs were counted in 3 independent experiments. Mean values ± SD are shown. (B) WT and Gal9 deficient NIH3T3 cells were stimulated with IFN-γ for 16 h and subsequently infected with T . gondii ME49 MOI10 for 24 and 48h. Genomic DNA from these samples was extracted and a T . gondii specific qPCR was performed. Parasite equivalents per 500 ng DNA are depicted for at least 10 individual samples from three independent experiments. Statistical analysis was performed using the Student’s t-test. ***: p < = 0.001; **: p < = 0.01; *: p < = 0.05.

Article Snippet: Additionally, WB analysis was performed with a polyclonal rabbit anti-mouse antibody against Gal9 (ARP54821_P050, Aviva Systems Biology).

Techniques: Clone Assay, CRISPR, Infection, Staining, Labeling, Confocal Microscopy

Journal: Scientific Reports

Article Title: Circulating galectin-9 as a novel prognosticator in patients with intrahepatic cholangiocarcinoma undergoing surgical resection

doi: 10.1038/s41598-025-15310-3

Figure Lengend Snippet: Tissue expression levels by IHC and prognostic value of GAL9 in resected specimens. A . IHC staining of GAL9 in surgical specimens of ICC. B . OS and RFS between the positive and negative groups of GAL9 IHC expression. C . OS and RFS between the positive and negative groups of GAL9 ( LGALS9 ) mRNA expression in the GAL9 IHC negative cohort (n = 40). D . Comparison of cGAL9 levels according to GAL9 mRNA positivity in the GAL9 IHC negative cohort. * Significant difference P < 0.05. Abbreviations: GAL9; galectin-9, IHC; immunohistochemistry, cGAL9; circulating galectin-9, ICC; intrahepatic cholangiocarcinoma, OS; overall survival, RFS; recurrence-free survival.

Article Snippet: The sections were incubated overnight at 4 °C with an anti-GAL9 primary antibody (Cell Signaling Technology, #54,330).

Techniques: Expressing, Immunohistochemistry, Comparison