Figure 7
Mechanistic studies for induced apoptosis via reactive oxygen species (ROS) generation.
(A) DNA laddering assay performed on genomic DNA extracted from MDA-MB231 cells (lane 1) untreated; or treated with (lane 2) C3-Lipocoat; (lane 3) C3 nanoparticles; (lane 4) CNP and (lane 5) β-carotene at concentration of 100 μM. Here lane 6 represent DNA ladder of 1–10 kb. Scatter plots obtained from 4% paraformaldehyde fixed MDA-MB231 cells (B) untreated or treated with (C) β-carotene; (D) C3 and (E) C3-Lipocoat at concentration of 100 μM post propidium iodide (PI; 10 μg/mL) staining. Scattering events in R7 and R2 gates represent the cells from live cell and apoptotic cell population, respectively. (F) Scattering events revealed the increasing % of apoptotic cell population in order of β-carotene, C3 and C3-Lipocoat while % live population followed the reverse order with maximum in untreated cells. (G) Variation in ROS abundance with time of incubation and change in β-carotene formulation in MDA-MB231 with maximum fold increase by C3-Lipocoat compared to CNP itself (H). Statistical analysis performed using Two-way ANOVA on IC50 values represented as *** for p < 0.001.
