Fig. 3

Effect of LDLc and HDLc supplementation on proliferation, cell cycle, and changes in signaling intermediate in colon cancer cells. HCT-116 cells were cultured in different concentrations of LDLc or HDLc, for different time points thereafter colony formation, cell cycle, and immunoblot analysis were performed as per the protocol mentioned in the methods. A Images of the colony were taken in a × 10 phase contrast microscope on the 12th day before staining with 0.05% crystal. B,C Images showing crystal violet-stained colonies of HCT-116 cells with or without LDLc, and HDLc treatment along with a bar graph representing the percent colony formed with respect to untreated controls. Colonies were stained with 0.05% crystal violet and images were captured in an Olympus DSLR camera and quantification was done using ImageJ software. Experiments were done in triplicate and performed twice. D,E Cell cycle analysis of HCT-116 cells treated with or without 50 µg/ml LDLc or HDLc for 48 h. D Histogram showing different phases of the cell cycle, and E bar graphs representing % cell population in different phases of the cell cycle, and the ratio of Go-G1/S-phase cell population with or without LDLc or HDLc treatment. Experiments were performed twice. F Immunoblot analysis of pERK and ERK protein expression was evaluated from the whole cell lysate of HCT-116 cells after treatment with different concentrations of LDLc or HDLc or vehicle control. Tubulin was used as a loading control. The experiment was performed twice