Journal: Oncotarget

Article Title: YBX1/YB-1 induces partial EMT and tumourigenicity through secretion of angiogenic factors into the extracellular microenvironment

doi:

Figure Lengend Snippet: a. Cell migration over 24 hr assessed by wound-healing assays and phase-contrast microscopy (4 × magnification). b. Cell migration as assessed by transwell assay using 8.0 μm membrane culture inserts. 24 h after cell seeding, migrating cells that passed through the transwell were stained with DAPI and counted ( n = 3; average ± SEM). c. Transwell invasion assays were conducted using 8.0 μm membrane inserts coated with 1 mg/mL Matrigel. 24 h after cell seeding, invading cells were stained with DAPI and counted ( n = 3; average ± SEM). d. Colony forming assays were performed in soft agar using CytoSelect 96-well cell transformation kit. 2,500 cells were inoculated and cultured for 7 days. Number of colonies were manually counted from 5 random fields of view (for each replicate) and colony size measured and quantified. Images are representative. (scale bar: 100 μm) ( n = 3; average ± SEM; ** P < 0.01).

Article Snippet: Cells were overlaid onto Transwell ® polycarbonate membrane cell culture inserts (8.0 μm pore size, Corning), and inserts placed into 24-well companion plates.

Techniques: Migration, Microscopy, Transwell Assay, Membrane, Staining, Transformation Assay, Cell Culture

Journal: Oncotarget

Article Title: YBX1/YB-1 induces partial EMT and tumourigenicity through secretion of angiogenic factors into the extracellular microenvironment

doi:

Figure Lengend Snippet: a. 5 × 10 4 2F-2B cells were conditioned with 30 μg of MDCK, MDCK YBX1 or 21D1 cell-derived secretome. Cell migration over 24 hr was assessed using Transwell inserts (8.0 μm pore size). Migrating cell nuclei were stained with DAPI, imaged and quantified ( n = 3; average ± SEM; ** P < 0.01). b. Cell-derived secretome was profiled using antibody-based angiogenesis arrays. Expression of 55 target proteins in the secretome was assessed. c. Relative expression of angiogenic modulators in cell-derived secretome ( n = 3; average intensity ± STD. * P < 0.05, ** P < 0.01).

Article Snippet: Cells were overlaid onto Transwell ® polycarbonate membrane cell culture inserts (8.0 μm pore size, Corning), and inserts placed into 24-well companion plates.

Techniques: Derivative Assay, Migration, Pore Size, Staining, Expressing

Journal: Nature immunology

Article Title: An αvβ3 integrin checkpoint is critical for efficient T H 2 cell cytokine polarization and potentiation of antigen-specific immunity.

doi: 10.1038/s41590-022-01378-w

Figure Lengend Snippet: Fig. 4 | IL-4/Gata3-mediated αvβ3 upregulation is required for TH2 cell differentiation in vitro. a, Flow cytometric analysis of αv and β3 expression by naive CD4+ T and differentiating TH1 and TH2 cells. d, Day. Data represent two independent experiments. b, Gata3 binding to Itgav and Itgb3 loci in TH2 cells as assessed by Gata3 ChIP–seq. c,d, Flow cytometric confirmation of retroviral Gata3 overexpression (c) and associated analyses of αv and β3 expression by transduced TH0 (neutral condition) cells (d). Data represent two independent experiments with four biologically independent samples in each experiment; mean ± s.d.; paired, two-sided Student’s t-test: *P = 0.0138, **P = 0.0073. e, Flow cytometric analysis of αv and β3 expression by ItgaviCD4KO TH cells cultured with

Article Snippet: Individual CRISPR sequences were inserted into the retroviral vector by ligation (New England Biolabs, T4 DNA ligase).

Techniques: Cell Differentiation, In Vitro, Expressing, Binding Assay, ChIP-sequencing, Retroviral, Over Expression, Cell Culture