Journal: ACS Nano

Article Title: Immune Modulatory Oxysterols Produced from Cholesterol-Containing Lipid Nanoparticles Regulate Tumor Growth

doi: 10.1021/acsnano.5c22020

Figure Lengend Snippet: LNP-oxysterols reduce tumor cell viability by inducing late apoptosis and necrosis in vitro. A) TC-1 tumor cell viability at 24, 48, and 72 h by MTT assay. B–E) Apoptosis and necrosis assessed at 24 h by propidium iodide (PI)/annexin V staining. Each data point represents a biological replicate. Statistical tests were performed by one-way ANOVA compared to vehicle without correction for multiple comparisons, where * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001; **** p ≤ 0.0001. F) Representative flow cytometry plots, where PI – /annexin V – are viable cells, PI + /annexin V + are cells in late apoptosis, PI – /annexin V + are cells in early apoptosis, and PI + /annexin V – are cells in necrosis. G) Pie charts of the percentage of viable, early apoptotic, late apoptotic, and necrotic cells.

Article Snippet: For proliferation studies, TC-1 tumor cells or BMDMs were plated in triplicate in 96-well plates, incubated at 37 °C with 5% CO 2 overnight for acclimation, and then treated with 55.7 μM LNP-cholesterol or LNP-oxysterols for 24, 48, and 72 h. Cytotoxicity and cell proliferation were assessed using the MTT assay kit (Cat. #4890050K, R&D Systems), and absorbance at 570 nm was measured using a Cytation 5 plate reader.

Techniques: In Vitro, MTT Assay, Staining, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Thrombospondin 1–CD47 Signalling Modulates Vascular Smooth Muscle Cell Senescence in Chronic Kidney Disease

doi: 10.3390/ijms27020755

Figure Lengend Snippet: TSP1 and indoxyl sulfate limit VSMC proliferation via CD47. hVSMC cell viability was measured by assessing reduction in tetrazolium salt sodium 3′- [1- [(phenylamino)-carbonyl]-3,4-tetrazolium]-bis(4-methoxy-6-nitro)benzene-sulfonic acid hydrate (XTT) and measuring absorbance at 450 nm in cells after 48 h treatment with ( A ) TSP1 (0, 0.2, 2.2, 5, 10 nM) ( n = 4), ( B ) TSP1 2.2 nM ± pre-treatment with anti-CD47 antibody for 30 min ( n = 6), ( C ) IS (0, 1, 10, 100, 500 µM) ( n = 4), or ( D ) IS (100, 500 µM) ± pre-treatment with anti-CD47 antibody for 30 min ( n = 4–8). All data shown are mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 by one-way analysis of variance with Holm–Sidak post-hoc test ( A , C ) or Kruskal–Wallis test ( B , D ). Abbreviations: αCD47—anti-CD47 antibody; hVSMC—human aortic vascular smooth muscle cell; IS—indoxyl sulfate; TSP1—thrombospondin-1.

Article Snippet: Cell proliferation and senescence were measured using XTT Cell Viability Kit (#9095, Cell Signalling Technology, Danvers, MA, USA) and Mammalian β-Galactosidase Assay Kit (75707, Thermo Fisher Scientific), respectively.

Techniques: