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