Journal: bioRxiv

Article Title: Cell-autonomous targeting of arabinogalactan by host immune factors inhibits mycobacterial growth

doi: 10.1101/2023.10.05.561003

Figure Lengend Snippet: Galectin-9 inhibits mycobacterial growth directly . A. Profile of Mtb H37Rv (Rv) grown at 37°C in Middlebrook 7H9 liquid medium with different concentration of Galectin-9 (Gal9, 0, 0.01, 0.1, 1, 10 μg/mL). Growth curve was measured using a Bioscreen Growth Curve Instrument. Optical density was measured at absorbance at 600 nm every 2 h. B. Growth profile of Mtb H37Rv (Rv) in Middlebrook 7H9 liquid medium with10μg/mL galectin-9 (Gal9) or inactivated galectin-9 (Gal9 HK, heat killed at 95 [for 5 min). C. CFU of Mtb H37Rv (Rv) on Middlebrook 7H10 solid medium with or without 10μg/mL Galectin-9 (Gal9). Cultures were grown at 37°C for 4-8 weeks. D. Growth profile of Mycobacterium smegmatis (MS) in Middlebrook 7H9 liquid medium with different concentrations of Galectin-9 (Gal9, 0, 0.01, 0.1, 1 μg/mL). E. Concentrations of galectin-9 in sera of healthy donors (n = 40) and active TB patients (n = 40). F. Confocal microscopy of M. bovis BCG-DsRed (BCG-DsRed, red) and Galectin[9 (Anti-Gal9, green) in THP-1 cells. Nuclei was stained with DAPI (blue). G. Percent of cells with galectin-9 positive (gal9 + ) BCG in total infected THP-1 cells. Symbols indicate colocalization ratio of at least 12 fields in each experiment. H. Confocal microscopy of Mtb H37Rv-GFP (Rv-GFP, green) and Galectin[9 (Anti-Gal9, red) in THP-1 cells. Nuclei were stained with DAPI (blue). I. Percent of cells with galectin9 positive (gal9 + ) Mtb H37Rv in total infected THP-1 cells. Symbols indicate colocalization ratio of at least 12 fields in each experiment. Data are shown as mean ± SD, n = 3 biologically independent experiments performed in triplicate (A-D). Data are representative of three independent experiments with similar results (F and H). Two-tailed unpaired Student’s t test (A-D, G, and I) or Mann-Whitney U test (E). P < 0.05 was considered statistically significant.

Article Snippet: Cells were stained with the anti-galectin-9 antibody (Cell Signaling Technology, Cat#54330, RRID:AB_2799456), antibodies at a dilution of 1:200 in 5% BSA in PBS overnight at 4[°C and then incubated with Alexa Fluor 488 or 555 conjugated secondary antibodies (Thermo Fisher Scientific, Cat# A-11008; RRID: AB_143165; Cat# A32732, RRID:AB_2633281) at a dilution of 1:1000 for 2[h at R.T..

Techniques: Concentration Assay, Confocal Microscopy, Staining, Infection, Two Tailed Test, MANN-WHITNEY

Journal: bioRxiv

Article Title: Cell-autonomous targeting of arabinogalactan by host immune factors inhibits mycobacterial growth

doi: 10.1101/2023.10.05.561003

Figure Lengend Snippet: A. Growth profile of Mtb H37Rv (Rv) in Middlebrook 7H9 liquid medium with or without galectin-9 (Gal9, 10 μg/mL) and lactose (1 μg/mL). B. Growth profile of Mtb H37Rv (Rv) in Middlebrook 7H9 liquid medium with or without galectin-9 (Gal9, 10 μg/mL) and D-glucose (10 μg/mL). C. Growth profile of Mtb H37Rv (Rv) in Middlebrook 7H9 liquid medium with or without galectin-9 (Gal9, 10 μg/mL) and AG (1 μg/mL). D. Growth profile of Mtb H37Rv (Rv) in Middlebrook 7H9 liquid medium with 1μg/mL CRD1 or CRD2 of galectin-9. Data are shown as mean ± SD, n = 3 biologically independent experiments performed in triplicate (A-D). Two-tailed unpaired Student’s t test (A-D). P < 0.05 was considered statistically significant.

Article Snippet: Cells were stained with the anti-galectin-9 antibody (Cell Signaling Technology, Cat#54330, RRID:AB_2799456), antibodies at a dilution of 1:200 in 5% BSA in PBS overnight at 4[°C and then incubated with Alexa Fluor 488 or 555 conjugated secondary antibodies (Thermo Fisher Scientific, Cat# A-11008; RRID: AB_143165; Cat# A32732, RRID:AB_2633281) at a dilution of 1:1000 for 2[h at R.T..

Techniques: Two Tailed Test

Journal: bioRxiv

Article Title: Cell-autonomous targeting of arabinogalactan by host immune factors inhibits mycobacterial growth

doi: 10.1101/2023.10.05.561003

Figure Lengend Snippet: A. Morphologic characteristics for Mtb H37Rv strain grown in liquid culture with or without anti-AG mAbs (1 μg/mL) observed by ×[2 magnifier. B. Bacterial shape of Mtb H37Rv strain treated as in (A) observed by acid fast staining under a Leica DM2500 microscope using the 100× oil microscopy. EMB, Ethambutol. Scale bar, 20μm. C. Ultrastructural morphology of Mtb H37Rv treated as in (A) analyzed by transmission electron microscopy (TEM). The cell wall was labeled with red arrows. D. Cell wall thickness of bacteria in (C). E. Schematic presentation of Mtb growth arrest by Galectin-9 or anti-AG antibodies. Data are representative of three independent experiments with similar results (A, B and C). Data are means ± SD of 11 bacteria, representatives of three independent experiments (D). Two-tailed unpaired Student’s t test (D). P < 0.05 was considered statistically significant.

Article Snippet: Cells were stained with the anti-galectin-9 antibody (Cell Signaling Technology, Cat#54330, RRID:AB_2799456), antibodies at a dilution of 1:200 in 5% BSA in PBS overnight at 4[°C and then incubated with Alexa Fluor 488 or 555 conjugated secondary antibodies (Thermo Fisher Scientific, Cat# A-11008; RRID: AB_143165; Cat# A32732, RRID:AB_2633281) at a dilution of 1:1000 for 2[h at R.T..

Techniques: Staining, Microscopy, Transmission Assay, Electron Microscopy, Labeling, Bacteria, Two Tailed Test

Journal: Journal of Cancer

Article Title: Galectin-9 and PSMB8 overexpression predict unfavorable prognosis in patients with AML

doi: 10.7150/jca.53686

Figure Lengend Snippet: LGALS9 was distinctively high expressed AML cell lines from CCLE analysis and prognostic significances of LGALS9 gene expression. (A) The mRNA expression level of LGALS9 ranked the second highest in a variety of cancer cell line (shown in red frame). (B) LGALS9 overexpression predicts unfavorable overall survival in patients with AML.

Article Snippet: Single-cell suspensions of mononuclear bone marrow cells were stained with the anti-galectin-9 antibody (BD Biosciences) and analyzed by flow cytometry using the Cell Quest software (FACSCalibur; BD Biosciences).

Techniques: Expressing, Over Expression

Journal: Journal of Cancer

Article Title: Galectin-9 and PSMB8 overexpression predict unfavorable prognosis in patients with AML

doi: 10.7150/jca.53686

Figure Lengend Snippet: KEGG and GO enrichment analysis revealing functional association of LGALS9 with immunoproteasome. (A) Venn diagram represents the intersection of the top 200 positively corrected genes between the UALCAN database and the GEPIA database, 138 common co-expressed genes were obtained. (B) GO enrichment of co-expressed genes in biological process, cellular component and molecular function. (C) KEGG enrichment analysis of co-expressed genes with LGALS9. False discovery rate ( FDR) < 0.05 was set as the cut-off criterion.

Article Snippet: Single-cell suspensions of mononuclear bone marrow cells were stained with the anti-galectin-9 antibody (BD Biosciences) and analyzed by flow cytometry using the Cell Quest software (FACSCalibur; BD Biosciences).

Techniques: Functional Assay

Journal: Journal of Cancer

Article Title: Galectin-9 and PSMB8 overexpression predict unfavorable prognosis in patients with AML

doi: 10.7150/jca.53686

Figure Lengend Snippet: Construction of PPI network of LGALS9 positive-correlation genes and analysis of hub genes. (A) Correlation between LGALS9 and PSMB8 mRNA expression determined using GEPIA database. (B) The top ten hub genes were identified using cytoHubba tool kits in Cytoscape. (C) Clustering analysis of LGALS9 co-expressed genes by STRING tools. (D) The KEGG pathway analysis of hub genes using WebGestalt web tool.

Article Snippet: Single-cell suspensions of mononuclear bone marrow cells were stained with the anti-galectin-9 antibody (BD Biosciences) and analyzed by flow cytometry using the Cell Quest software (FACSCalibur; BD Biosciences).

Techniques: Expressing