Journal: Science Advances

Article Title: Galectin-3 exacerbates autoimmune diabetes by limiting regulatory T cell differentiation and function

doi: 10.1126/sciadv.adz7916

Figure Lengend Snippet: ( A ) The concentration of fasting serum Galectin-3 in healthy controls (HC, n = 132), Ab − FDR ( n = 76), Ab + FDR ( n = 30), and patients with T1D ( n = 234). Data are expressed as median ± range. ( B ) The mRNA expression levels of Galectin-3 in CD14 + monocytes isolated from patients with T1D and HC ( n = 10), calculated using the 2 −△△Ct method. ( C ) The concentration of fasting serum LPS in HC and T1D groups ( n = 56). Data are expressed as median ± range. ( D ) Correlation between serum levels of Galectin-3 and LPS in HC and patients with T1D. ( E to J ) The THP-1 cell line, THP-1–cell induced macrophages, and the RAW 264.7 cell line were treated with LPS (100 ng/ml) or vehicle for 24 hours. The mRNA expression levels of Galectin-3 [(E), (G), and (I)] and the protein concentration of Galectin-3 in the supernatant [(F), (H), and (J)] were measured. Data in (E) to (J) are representative of at least three independent experiments with similar results. Data are expressed as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

Article Snippet: Subsequently, cells were washed with phosphate-buffered saline (PBS), followed by isolation of monocytes from PBMCs using anti-human CD14 immunomagnetic beads (Miltenyi Biotec, Germany).

Techniques: Concentration Assay, Expressing, Isolation, Protein Concentration

Journal: PLOS One

Article Title: UTX/Top2β axis mediated spinal cord microvascular endothelial cells senescence exacerbates spinal cord injury

doi: 10.1371/journal.pone.0338326

Figure Lengend Snippet: Representative immunofluorescence images and quantification of CD31 + (green), UTX + (red) or p16 INK4a + (red), and DAPI (blue) in the injured spinal cord of mice at sham, 3, 5, 7, and 14 days post-SCI. Data are mean ± SEM; Scale bars: 50 μm. *p < 0.05, **p < 0.01, ****p < 0.0001 vs. sham. NS, not significant. SCMVECs, spinal cord microvascular endothelial cells; SCI, spinal cord injury.

Article Snippet: CD31 + cells were isolated utilizing CD31 immunomagnetic beads (Miltenyi Biotec, 130-097-418) according to the instruction provided by the manufacturer.

Techniques: Immunofluorescence

Journal: PLOS One

Article Title: UTX/Top2β axis mediated spinal cord microvascular endothelial cells senescence exacerbates spinal cord injury

doi: 10.1371/journal.pone.0338326

Figure Lengend Snippet: (A) Schematic of transgenic mice generation. (B) CD31 + (green), UTX + (red), and DAPI (blue) staining in spinal cords of UTX -/- and UTX f/f mice. (C) CD31 + (green), p16 INK4a (red), and DAPI (blue) staining in the spinal cord injury region of UTX -/- and UTX f/f mice at 7 day post SCI. Data are mean ± SEM; Scale bars: 50 μm. *p < 0.05 vs. UTX f/f . SCI, spinal cord injury.

Article Snippet: CD31 + cells were isolated utilizing CD31 immunomagnetic beads (Miltenyi Biotec, 130-097-418) according to the instruction provided by the manufacturer.

Techniques: Transgenic Assay, Staining

Journal: PLOS One

Article Title: UTX/Top2β axis mediated spinal cord microvascular endothelial cells senescence exacerbates spinal cord injury

doi: 10.1371/journal.pone.0338326

Figure Lengend Snippet: (A) Phase-contrast images of primary SCMVECs from UTX f/f and UTX -/- mice. (B) Flow cytometry showing >90% CD31 + purity in UTX f/f mice. (C) qRT-PCR analysis of UTX mRNA from UTX f/f and UTX -/- mice. (D) Representative images and quantitative analyses of CD31 + (green), UTX + (red), and DAPI (blue) in cultured primary SCMVECs from UTX f/f and UTX -/- mice. Data are mean ± SEM. Scale bars: 50 μm. *p < 0.05, **p < 0.01 vs. UTX f/f .

Article Snippet: CD31 + cells were isolated utilizing CD31 immunomagnetic beads (Miltenyi Biotec, 130-097-418) according to the instruction provided by the manufacturer.

Techniques: Flow Cytometry, Quantitative RT-PCR, Cell Culture

Journal: PLOS One

Article Title: UTX/Top2β axis mediated spinal cord microvascular endothelial cells senescence exacerbates spinal cord injury

doi: 10.1371/journal.pone.0338326

Figure Lengend Snippet: Representative images and quantification of CD31 + , p16 INK4a (red), and DAPI (blue) (A); immunoblotting of p16 INK4a and UTX (B); SA-β-gal staining (C); CCK-8 cell viability assay (D); and CD31 + , Ki67 + (red), and DAPI (blue) staining (E) in primary SCMVECs from UTX f/f and UTX -/- mice. Data are mean ± SEM; scale bars: 50 μm. *p < 0.05, **p < 0.01 vs. UTX f/f group. SCMVECs, spinal cord microvascular endothelial cells; SCI, spinal cord injury.

Article Snippet: CD31 + cells were isolated utilizing CD31 immunomagnetic beads (Miltenyi Biotec, 130-097-418) according to the instruction provided by the manufacturer.

Techniques: Western Blot, Staining, CCK-8 Assay, Viability Assay

Journal: PLOS One

Article Title: UTX/Top2β axis mediated spinal cord microvascular endothelial cells senescence exacerbates spinal cord injury

doi: 10.1371/journal.pone.0338326

Figure Lengend Snippet: UTX deletion reduced senescence and promoted vascular regeneration by upregulating Top2β. (A) qRT-PCR analysis of Top2β mRNA; (B) Immunoblotting of Top2β protein; (C) Immunofluorescence of CD31 + (green), Top2β + (red), and DAPI (blue) nuclei in UTX f/f and UTX -/- SCMVECs. (D)Relative Top2β mRNA expression in siNC vs. siTop2β groups by qRT-PCR. (E) Representative immunoblots and quantitative analysis of p16 INK4a protein in indicated groups. (F) Cell viability of SCMVECs in indicated groups. Data represent means ± SEM; *p < 0.05, **p < 0.01 vs. UTX f/f group.

Article Snippet: CD31 + cells were isolated utilizing CD31 immunomagnetic beads (Miltenyi Biotec, 130-097-418) according to the instruction provided by the manufacturer.

Techniques: Quantitative RT-PCR, Western Blot, Immunofluorescence, Expressing